{"gene":"CRY2","run_date":"2026-06-09T22:57:19","timeline":{"discoveries":[{"year":1999,"finding":"Mice lacking both CRY1 and CRY2 show instantaneous and complete loss of free-running circadian rhythmicity, demonstrating that CRY1 and CRY2 are essential for maintenance of circadian rhythms (not merely photoreceptors). CRY1-null mice show accelerated and CRY2-null mice show delayed free-running locomotor activity periodicity.","method":"Genetic knockout (Cry1-/-, Cry2-/-, Cry1-/-/Cry2-/- mice) with locomotor activity monitoring","journal":"Nature","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean knockout with defined behavioral phenotype, replicated across multiple genotypes in a landmark study","pmids":["10217146"],"is_preprint":false},{"year":2002,"finding":"CRY2 (and CRY1, PER2) activates transcription of the mouse Bmal1 gene, while BMAL1-CLOCK dimers repress it, establishing an interlocked feedback loop within the circadian clockwork.","method":"Transcriptional reporter assays in cell culture; promoter characterization of mBmal1","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — cell-based reporter assays, single lab, but consistent with broader circadian clock model","pmids":["11798163"],"is_preprint":false},{"year":2001,"finding":"An intact flavin binding domain is required for CRY2 function in suppressing CLOCK/BMAL1-mediated transcription. Mutation of any of the three conserved tryptophan residues in the putative electron transport chain inhibits xCRY2b function, indicating CRY2 requires electron transport through the conserved tryptophan pathway, unlike CRY1 which depends only on the last tryptophan.","method":"Site-directed mutagenesis of conserved tryptophan residues; transcriptional suppression assays in Xenopus CRY2","journal":"Current biology","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — mutagenesis with functional readout, but single lab using Xenopus ortholog, not mammalian CRY2","pmids":["11747820"],"is_preprint":false},{"year":2010,"finding":"CRY2 undergoes proteasomal degradation initiated by a dual-kinase mechanism: DYRK1A phosphorylates Ser557 as a priming event, enabling subsequent GSK-3β-mediated phosphorylation of Ser553, which leads to proteasomal degradation of CRY2. Knockdown of Dyrk1a causes abnormal cytosolic CRY2 accumulation and shortens circadian period.","method":"In vitro kinase assays, site-directed mutagenesis (S557A/S553A), RNAi knockdown in cells, circadian period measurement","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro kinase assay, mutagenesis, and cellular phenotype from single lab with multiple orthogonal methods","pmids":["20123978"],"is_preprint":false},{"year":2010,"finding":"Two residues in the C-terminal domain of mouse CRY2, Arg-501 and Lys-503 (within residues 493–512), are essential for direct physical interaction with PER2. Mutation of both residues abolishes CRY2-PER2 interaction.","method":"Mammalian two-hybrid assay, co-immunoprecipitation, oligonucleotide-based degenerate PCR mutagenesis","journal":"BMC molecular biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP and two-hybrid with mutagenesis, single lab","pmids":["20840750"],"is_preprint":false},{"year":2013,"finding":"CRY1 and CRY2 are both transcriptional repressors within the SCN clockwork, but CRY1 is significantly more potent than CRY2. CRY1 prolongs the interval of transcriptional suppression while CRY2 does not, and CRY2 attenuates the period-lengthening effects of CRY1. Both CRYs dose-dependently lengthen the intrinsic high-frequency SCN rhythm.","method":"Genetic epistasis using Fbxl3(Afh) allele in Cry1- and Cry2-deficient mouse backgrounds; SCN bioluminescence recording; wheel-running analysis","journal":"The Journal of neuroscience","confidence":"High","confidence_rationale":"Tier 2 / Strong — epistasis analysis across multiple genotypes with two independent phenotypic readouts (behavior and bioluminescence)","pmids":["23616524"],"is_preprint":false},{"year":2009,"finding":"Random mutagenesis identified CRY2-specific residues required for repression of CLOCK-BMAL1. CRY2(G354D) is deficient in clock protein binding required for repression by both CRYs, while CRY2(G351D) shows normal binding but a CRY2-specific repression defect. Overexpression of CRY2(G351D) abolishes circadian rhythmicity, implicating a CRY2-unique repression mechanism.","method":"Random mutagenesis screen, cell-based CLOCK-BMAL1 reporter assay, co-immunoprecipitation, circadian rhythm analysis in NIH 3T3 cells","journal":"Molecular and cellular biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mutagenesis screen with functional readout and binding assays, single lab","pmids":["19687303"],"is_preprint":false},{"year":2015,"finding":"FBXW7 is an E3 ubiquitin ligase that targets CRY2 for proteasomal degradation by binding directly to phosphorylated Thr300 of CRY2, enhancing CRY2 ubiquitination and accelerating CRY2 turnover.","method":"Co-immunoprecipitation, ubiquitination assays, site-directed mutagenesis (Thr300), cycloheximide chase; biochemical and cell biology analyses","journal":"Molecular cancer therapeutics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct binding and ubiquitination assays with phospho-site mutagenesis, single lab","pmids":["25855785"],"is_preprint":false},{"year":2016,"finding":"CRY2 functions as a component of an FBXL3-containing SCF E3 ubiquitin ligase complex that recruits T58-phosphorylated c-MYC for ubiquitylation and degradation. CRY1 cannot substitute for CRY2 in this function. This represents a circadian mechanism for control of cell proliferation via c-MYC turnover.","method":"Co-immunoprecipitation, ubiquitylation assays, phospho-specific recruitment assays; genetic complementation showing CRY1 cannot substitute","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, in-cell ubiquitination assays, isoform specificity established, multiple orthogonal approaches","pmids":["27840026"],"is_preprint":false},{"year":2017,"finding":"CRY1 and CRY2 broadly interact with nuclear hormone receptors and serve as corepressors for many NRs, binding independently of other core clock factors to genomic sites enriched for NR recognition motifs, contributing to circadian regulation of drug metabolism.","method":"Genomic binding analysis (ChIP-related), co-repressor functional assays; interaction studies with multiple NRs","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genomic binding and functional corepressor assays, single lab with multiple NRs tested","pmids":["28751364"],"is_preprint":false},{"year":2018,"finding":"CRY2, but not CRY1, specifically interacts with Bclaf1 to stabilize mRNAs encoding cyclin D1 and Tmem176b, regulating circadian patterns of myoblast proliferation and myogenic cell fusion. Cry2-/- myoblasts show premature cell cycle exit and form short myotubes, and muscle regeneration is impaired in Cry2-/- mice.","method":"Co-immunoprecipitation of CRY2-Bclaf1 complex, Cry2-/- mouse model, mRNA stability assays, single myofiber analysis, muscle regeneration assays","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, knockout mouse with defined phenotype, mRNA stability assays, Bclaf1 knockdown phenocopy, multiple orthogonal methods","pmids":["29466738"],"is_preprint":false},{"year":2021,"finding":"Two cancer-associated CRY2 missense mutations (D325H and S510L in mouse) suppress P53 target-gene expression and accelerate growth of primary fibroblasts expressing high c-MYC. The mutations have divergent impacts on circadian rhythms and on CRY2's ability to interact with SCF-FBXL3, and neither affects steady-state levels of overexpressed c-MYC.","method":"Stable expression of CRY2 mutants in primary mouse fibroblasts, P53 target gene expression analysis, circadian rhythm assays, co-immunoprecipitation with FBXL3","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — cell-based functional assays with defined mutants, Co-IP, single lab with multiple readouts","pmids":["34183418"],"is_preprint":false},{"year":2022,"finding":"X-ray crystal structure of mammalian CRY2 in complex with the small molecule SHP656 reveals that compound binding is compatible with the intrinsic CRY2 gatekeeper W417 'in' orientation. The gatekeeper residue W417 and lid loop interactions are important for CRY2 isoform selectivity of this compound. SHP656 lengthened cellular circadian period in a CRY2-dependent manner.","method":"X-ray crystallography, molecular dynamics simulations, circadian period assays in CRY2-dependent manner, mutagenesis of W417","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure with mutagenesis validation and cellular functional assay, single lab","pmids":["36161947"],"is_preprint":false},{"year":2019,"finding":"Human CRY2 knockout cells generated by CRISPR/Cas9 show long-period circadian rhythms, while CRY1/CRY2 double knockout cells are arrhythmic, consistent with mouse knockout models and confirming CRY2's period-setting role in human cells.","method":"CRISPR/Cas9 knockout of CRY2 in human U-2 OS cells; circadian bioluminescence recording","journal":"Frontiers in physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean CRISPR KO with defined circadian phenotype, single lab, consistent with prior mouse data","pmids":["31143130"],"is_preprint":false},{"year":2023,"finding":"CRY2 variant p.Ser420Phe has reduced affinity for PER2 and defective nuclear translocation, resulting in reduced repression of CLOCK:BMAL1-driven transcription. This variant is unexpectedly resistant to degradation via FBXL3 and FBXL21 (canonical proteasomal pathway), indicating Ser-420 is required for E3 ligase interaction. Wild-type CRY2 (and this variant) can also be degraded via a lysosomal pathway. The variant causes a ~7 h shorter circadian period in Cry1-/-Cry2-/- MEFs.","method":"Site-directed mutagenesis, co-immunoprecipitation with PER2/FBXL3/FBXL21, nuclear localization imaging, proteasomal/lysosomal inhibitor experiments, complementation in Cry1-/-Cry2-/- double KO MEFs","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — mutagenesis, multiple Co-IP experiments, degradation pathway dissection with inhibitors, functional complementation, single lab with multiple orthogonal methods","pmids":["37951306"],"is_preprint":false},{"year":2023,"finding":"CRY2 represses CLOCK/BMAL1-induced transcription via a critical Cys432 residue that mediates PER2 heterodimer formation. The C432 mutation disrupts PER2 association without affecting BMAL1 binding. This CRY2-PER2 repression complex is required for transcriptional repression of Wnt pathway components to promote adipogenesis.","method":"Site-directed mutagenesis (C432), co-immunoprecipitation, transcriptional reporter assays, adipogenic differentiation assays, CRY2 KD/OE in preadipocytes, KL001 stabilization","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — mutagenesis identifying specific residue, Co-IP, functional adipogenesis assay, multiple orthogonal methods in single lab","pmids":["37724597"],"is_preprint":false},{"year":2020,"finding":"FBXW7 promotes CRY2 ubiquitin-mediated degradation and thereby promotes trophoblast migration and invasion. The lncRNA MALAT1 recruits FBXW7 to impair CRY2 protein stability; reduced MALAT1 leads to CRY2 accumulation and suppressed migration/invasion.","method":"RNA pull-down, co-immunoprecipitation of FBXW7-CRY2, CRY2 protein stability assays, trophoblast migration/invasion assays","journal":"Journal of cellular physiology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — Co-IP and functional assays, single lab, consistent with FBXW7-CRY2 interaction from prior work","pmids":["32776544"],"is_preprint":false},{"year":2020,"finding":"CRY2 overexpression impairs trophoblast migration and invasion by inhibiting the c-Myc-BMAL1 pathway. c-Myc binds the BMAL1 promoter to induce BMAL1 transcription, which activates MMP2/9. CRY2 suppresses this c-Myc-BMAL1-MMP2/9 axis.","method":"Luciferase reporter assay, chromatin immunoprecipitation (c-Myc on BMAL1 promoter), wound healing and Transwell invasion assays, CRY2 overexpression/knockdown in HTR-8/SVneo cells","journal":"Journal of biochemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP, reporter assay, and functional migration assays with CRY2 OE/KD, single lab","pmids":["31536114"],"is_preprint":false},{"year":2023,"finding":"Loss of CRY2 in satellite cells enhances muscle regeneration by activating ERK1/2 signaling and ETS1, which binds the PAX7 promoter to induce PAX7 transcription and increase satellite cell proliferation.","method":"Satellite cell/skeletal muscle lineage-specific Cry2 knockout mice (CRY2scko), immunostaining, single myofiber analysis, ERK1/2 activation assays, ChIP (ETS1 on PAX7 promoter)","journal":"MedComm","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — tissue-specific KO mouse, ChIP, signaling pathway characterization, single lab","pmids":["36636367"],"is_preprint":false},{"year":2025,"finding":"CRY2 physically interacts with TRPC1, and this interaction is detected by co-immunoprecipitation. Both proteins co-translocate to the nucleus following pulsed electromagnetic field (PEMF) exposure. CRY2 overexpression enhances PEMF-induced myogenic responses, while CRY2 silencing reduces them. Reducing FAD content by silencing riboflavin kinase attenuates PEMF responsiveness.","method":"Co-immunoprecipitation, immunofluorescence co-localization, CRY2 overexpression/knockdown, riboflavin kinase silencing, PEMF exposure assays in myoblasts","journal":"Cells","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — Co-IP and co-localization with functional OE/KD assays, single lab, novel interaction","pmids":["39937022"],"is_preprint":false},{"year":2025,"finding":"CRY2 plays a central role in serum-induced circadian clock phase resetting. Steroid hormone receptors (including sex-hormone receptors) are key mediators of serum-induced phase resetting, and CRY2 mediates their effect on clock phase independently of its role in period-length determination.","method":"Circa-SCOPE high-throughput single-cell phase transition curve analysis; CRY2 perturbation experiments; steroid hormone receptor pathway analysis","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — novel high-throughput single-cell method with CRY2 perturbation, single lab","pmids":["41354650"],"is_preprint":false},{"year":2025,"finding":"CRY2 overexpression in mouse renal tubular cells alleviates cisplatin-induced cytotoxicity by reducing platinum accumulation through upregulation of efflux transporters ATP7A and MRP2.","method":"CRY2 overexpression in MuRTE61 cells, cell viability assays, platinum content measurement, western blot of ATP7A and MRP2","journal":"Biological & pharmaceutical bulletin","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single OE experiment with defined readout, single lab, single method for mechanism","pmids":["40254427"],"is_preprint":false},{"year":2026,"finding":"CRY2 nuclear-cytoplasmic shuttling in chondrocytes depends on ROCK activation and actin polymerization under mechanical loading. Inhibition of ROCK causes actin depolymerization and partially blocks CRY2 nuclear-cytoplasmic trafficking. Knockdown of CRY2 attenuates mechanical loading-sustained circadian oscillations and cartilage homeostasis.","method":"ROCK inhibitor treatment, actin polymerization assays, CRY2 localization imaging (nuclear-cytoplasmic shuttling), CRY2 knockdown in chondrocytes and condyle explants","journal":"Bone & joint research","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — direct localization with mechanistic inhibitor experiments and functional KD readout, single lab","pmids":["41490462"],"is_preprint":false}],"current_model":"Mammalian CRY2 is a core circadian transcriptional repressor that suppresses CLOCK/BMAL1-driven transcription via a PER2-interacting C-terminal domain (requiring residues including Arg-501, Lys-503, and Cys-432), undergoes circadian-regulated proteasomal degradation initiated by DYRK1A-mediated priming phosphorylation at Ser557 followed by GSK-3β phosphorylation at Ser553 (with FBXL3 and FBXL21 as E3 ligases and a secondary lysosomal degradation pathway), and performs non-redundant clock-output functions including acting as an adaptor within SCF-FBXL3 to recruit phospho-T58 c-MYC for ubiquitylation, interacting with Bclaf1 to stabilize myogenic mRNAs, serving as a corepressor for nuclear hormone receptors, and mediating serum-induced phase resetting via steroid hormone receptors."},"narrative":{"mechanistic_narrative":"CRY2 is a core component of the mammalian circadian clockwork that acts as a transcriptional repressor of the CLOCK/BMAL1 complex, and its loss alters or, in combination with CRY1, abolishes free-running circadian rhythmicity [PMID:10217146, PMID:31143130]. Repression depends on a C-terminal domain that mediates direct heterodimerization with PER2 through residues including Arg-501/Lys-503 and Cys-432, the latter disrupting PER2 association without affecting BMAL1 binding, and on additional residues such as Gly-351 that define a CRY2-specific repression activity distinct from CRY1 [PMID:20840750, PMID:37724597, PMID:19687303]; an intact flavin/electron-transport pathway is also required for CLOCK/BMAL1 suppression [PMID:11747820]. CRY2 abundance is set by regulated proteolysis: DYRK1A primes Ser557 phosphorylation to enable GSK-3β phosphorylation of Ser553 and proteasomal turnover, while FBXL3/FBXL21 and FBXW7 (the latter binding phospho-Thr300) target CRY2 for ubiquitin-mediated degradation, with Ser-420 required for E3 ligase engagement and a parallel lysosomal route also operating [PMID:20123978, PMID:25855785, PMID:37951306]. Beyond the canonical loop, CRY2 performs non-redundant, isoform-specific output functions that CRY1 cannot substitute: it serves as an adaptor in an SCF-FBXL3 complex to recruit phospho-T58 c-MYC for ubiquitylation, links the clock to cell proliferation and P53 target regulation, and binds Bclaf1 to stabilize myogenic mRNAs controlling myoblast proliferation and muscle regeneration [PMID:27840026, PMID:34183418, PMID:29466738]. CRY2 additionally acts as a corepressor for nuclear hormone receptors at NR-motif-enriched genomic sites and mediates steroid-hormone-receptor-dependent serum-induced phase resetting independently of its period-setting role [PMID:28751364, PMID:41354650]. A crystal structure of CRY2 bound to the period-lengthening compound SHP656 defines a gatekeeper W417 conformation underlying isoform-selective small-molecule modulation [PMID:36161947].","teleology":[{"year":1999,"claim":"Established that CRY2 is not merely a photoreceptor but an essential clock component, resolving whether cryptochromes maintain mammalian rhythmicity.","evidence":"Single and double Cry1/Cry2 knockout mice with locomotor activity monitoring","pmids":["10217146"],"confidence":"High","gaps":["Did not define the molecular mechanism of repression","Behavioral phenotype does not localize CRY2 action to specific molecular partners"]},{"year":2001,"claim":"Addressed whether the flavin/electron-transport apparatus is functionally required for transcriptional repression, distinguishing CRY2 from CRY1.","evidence":"Site-directed mutagenesis of conserved tryptophans with transcriptional suppression assays in Xenopus CRY2","pmids":["11747820"],"confidence":"Medium","gaps":["Used Xenopus ortholog, not mammalian CRY2","Did not establish a photochemical or redox mechanism in mammalian cells"]},{"year":2002,"claim":"Defined the interlocked feedback architecture by showing CRY2 activates Bmal1 transcription while CLOCK/BMAL1 represses it.","evidence":"Transcriptional reporter assays and mBmal1 promoter characterization in cell culture","pmids":["11798163"],"confidence":"Medium","gaps":["Reporter-based; in vivo contribution to the loop not directly tested","Mechanism of CRY2-mediated activation unresolved"]},{"year":2009,"claim":"Distinguished clock-protein binding from a CRY2-unique repression step, showing CRY2 has a repression mechanism separable from complex assembly.","evidence":"Random mutagenesis screen (G354D, G351D), reporter assays, Co-IP and rhythm analysis in NIH 3T3 cells","pmids":["19687303"],"confidence":"Medium","gaps":["Molecular nature of the CRY2-specific repression defect not defined","Single lab, cell-based"]},{"year":2010,"claim":"Mapped the PER2 interaction surface and the priming-phosphorylation degradation switch, defining how CRY2 abundance is timed.","evidence":"Two-hybrid/Co-IP mutagenesis (R501/K503) and in vitro DYRK1A/GSK-3β kinase assays with S557A/S553A mutants and RNAi","pmids":["20840750","20123978"],"confidence":"High","gaps":["Did not identify the cognate E3 ligase acting on phospho-CRY2","Kinase priming hierarchy shown in vitro and cells but not in vivo timing"]},{"year":2013,"claim":"Quantified the functional asymmetry between CRY1 and CRY2, showing CRY2 is a weaker repressor that modulates CRY1 action.","evidence":"Fbxl3(Afh) epistasis across Cry-deficient backgrounds with SCN bioluminescence and wheel-running","pmids":["23616524"],"confidence":"High","gaps":["Structural basis of differential repressor potency not resolved","Does not explain CRY2's non-redundant output roles"]},{"year":2015,"claim":"Identified FBXW7 as a phospho-Thr300-dependent E3 ligase for CRY2, expanding the degradation machinery beyond FBXL3/FBXL21.","evidence":"Co-IP, ubiquitination assays, Thr300 mutagenesis and cycloheximide chase","pmids":["25855785"],"confidence":"Medium","gaps":["Kinase phosphorylating Thr300 not identified","Relative contribution versus FBXL3/FBXL21 unresolved"]},{"year":2016,"claim":"Revealed a non-redundant output function: CRY2 acts as an SCF-FBXL3 adaptor recruiting phospho-T58 c-MYC, linking the clock to proliferation.","evidence":"Co-IP, in-cell ubiquitylation, phospho-specific recruitment and CRY1 non-complementation","pmids":["27840026"],"confidence":"High","gaps":["In vivo tumor-suppressive role not established here","Structural basis of the c-MYC recruitment specificity unknown"]},{"year":2017,"claim":"Showed CRY2 broadly corepresses nuclear hormone receptors independently of core clock factors, connecting the clock to drug metabolism.","evidence":"Genomic binding analysis and corepressor functional assays across multiple NRs","pmids":["28751364"],"confidence":"Medium","gaps":["CRY2-versus-CRY1 specificity at NR sites not dissected","Direct versus indirect genomic recruitment unclear"]},{"year":2018,"claim":"Demonstrated a post-transcriptional output role: CRY2-specific binding to Bclaf1 stabilizes myogenic mRNAs and controls muscle regeneration.","evidence":"Co-IP, Cry2-/- mice, mRNA stability assays, single myofiber and regeneration assays","pmids":["29466738"],"confidence":"High","gaps":["Mechanism of CRY2/Bclaf1 mRNA selectivity unknown","Relationship to CRY2's nuclear repressor role not integrated"]},{"year":2019,"claim":"Confirmed CRY2's period-setting role in human cells, validating cross-species conservation of the phenotype.","evidence":"CRISPR/Cas9 CRY2 and CRY1/CRY2 knockouts in U-2 OS cells with bioluminescence","pmids":["31143130"],"confidence":"Medium","gaps":["No molecular dissection beyond period phenotype","Output functions not tested in this system"]},{"year":2020,"claim":"Connected CRY2 turnover to trophoblast behavior, showing FBXW7-driven degradation and CRY2 suppression of the c-Myc-BMAL1-MMP axis.","evidence":"RNA pull-down, Co-IP, stability and migration/invasion assays; ChIP of c-Myc on BMAL1 promoter in HTR-8/SVneo cells","pmids":["32776544","31536114"],"confidence":"Medium","gaps":["In vivo relevance to placentation not established","Single cell-line systems"]},{"year":2021,"claim":"Linked cancer-associated CRY2 missense variants to P53 target suppression and c-MYC-dependent growth, implicating CRY2 in tumor biology.","evidence":"Stable CRY2 mutant expression in primary fibroblasts, P53 target and circadian assays, FBXL3 Co-IP","pmids":["34183418"],"confidence":"Medium","gaps":["Mechanism connecting variants to P53 targets undefined","No in vivo tumorigenesis data"]},{"year":2022,"claim":"Provided a structural basis for isoform-selective small-molecule modulation, defining the W417 gatekeeper conformation.","evidence":"X-ray crystallography of CRY2-SHP656, MD simulations, W417 mutagenesis and CRY2-dependent period assays","pmids":["36161947"],"confidence":"High","gaps":["Structure of full repressor complex with PER2/CLOCK/BMAL1 not resolved","Endogenous ligand state not addressed"]},{"year":2023,"claim":"Resolved the Cys432/Ser420 determinants of PER2 binding, repression and E3 engagement, and extended CRY2 repression to Wnt-driven adipogenesis and ERK/ETS1-PAX7 muscle programs.","evidence":"C432 and S420F mutagenesis with PER2/FBXL3/FBXL21 Co-IP, localization imaging, degradation inhibitors, adipogenesis assays, and satellite-cell Cry2 KO with ChIP","pmids":["37724597","37951306","36636367"],"confidence":"High","gaps":["How a single residue couples binding, localization and degradation not fully mechanized","Tissue-specific output mechanisms not unified"]},{"year":2025,"claim":"Identified period-independent and signaling-coupled roles: CRY2 mediates steroid-hormone-receptor-dependent phase resetting, interacts with TRPC1 in electromagnetic-field responses, and modulates renal drug efflux.","evidence":"Single-cell phase-transition (Circa-SCOPE) with CRY2 perturbation; Co-IP/co-localization with TRPC1 and riboflavin-kinase silencing in myoblasts; CRY2 overexpression with ATP7A/MRP2 readouts in renal cells","pmids":["41354650","39937022","40254427"],"confidence":"Medium","gaps":["TRPC1 interaction is single-lab without reciprocal in vivo validation","Renal efflux role rests on a single overexpression experiment (Low confidence)","Molecular link between steroid receptors and CRY2 phase resetting undefined"]},{"year":2026,"claim":"Showed CRY2 nuclear-cytoplasmic shuttling is mechanosensitive, coupling ROCK/actin dynamics to clock oscillations in cartilage.","evidence":"ROCK inhibition, actin polymerization assays, CRY2 localization imaging and knockdown in chondrocytes and condyle explants","pmids":["41490462"],"confidence":"Medium","gaps":["Direct molecular link between actin and CRY2 transport unknown","In vivo cartilage relevance from explant model only"]},{"year":null,"claim":"How CRY2's distinct activities — transcriptional repression, E3-adaptor function, mRNA stabilization, NR corepression, and mechano-/hormone-coupled localization — are integrated and isoform-specifically partitioned from CRY1 remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structure of the assembled CRY2-PER2-CLOCK/BMAL1 repressor complex","Mechanism determining when CRY2 acts as repressor versus SCF adaptor versus RNA-stabilizing factor unknown","Endogenous flavin/redox cofactor function in mammalian CRY2 not established"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[0,1,6,9,15]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[8,11]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[8,10]},{"term_id":"GO:0003723","term_label":"RNA binding","supporting_discovery_ids":[10]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[9,15]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[14,22]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[3,22]},{"term_id":"GO:0005654","term_label":"nucleoplasm","supporting_discovery_ids":[9,15]}],"pathway":[{"term_id":"R-HSA-9909396","term_label":"Circadian clock","supporting_discovery_ids":[0,5,13,20]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[1,6,9,15]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[3,7,8,14]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[8,10,11]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[10,15,18]}],"complexes":["CRY2-PER2 repressor complex","SCF-FBXL3 E3 ubiquitin ligase complex"],"partners":["PER2","BMAL1","CLOCK","FBXL3","FBXW7","C-MYC","BCLAF1","TRPC1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q49AN0","full_name":"Cryptochrome-2","aliases":[],"length_aa":593,"mass_kda":66.9,"function":"Transcriptional repressor which forms a core component of the circadian clock. The circadian clock, an internal time-keeping system, regulates various physiological processes through the generation of approximately 24 hour circadian rhythms in gene expression, which are translated into rhythms in metabolism and behavior. It is derived from the Latin roots 'circa' (about) and 'diem' (day) and acts as an important regulator of a wide array of physiological functions including metabolism, sleep, body temperature, blood pressure, endocrine, immune, cardiovascular, and renal function. Consists of two major components: the central clock, residing in the suprachiasmatic nucleus (SCN) of the brain, and the peripheral clocks that are present in nearly every tissue and organ system. Both the central and peripheral clocks can be reset by environmental cues, also known as Zeitgebers (German for 'timegivers'). The predominant Zeitgeber for the central clock is light, which is sensed by retina and signals directly to the SCN. The central clock entrains the peripheral clocks through neuronal and hormonal signals, body temperature and feeding-related cues, aligning all clocks with the external light/dark cycle. Circadian rhythms allow an organism to achieve temporal homeostasis with its environment at the molecular level by regulating gene expression to create a peak of protein expression once every 24 hours to control when a particular physiological process is most active with respect to the solar day. Transcription and translation of core clock components (CLOCK, NPAS2, BMAL1, BMAL2, PER1, PER2, PER3, CRY1 and CRY2) plays a critical role in rhythm generation, whereas delays imposed by post-translational modifications (PTMs) are important for determining the period (tau) of the rhythms (tau refers to the period of a rhythm and is the length, in time, of one complete cycle). A diurnal rhythm is synchronized with the day/night cycle, while the ultradian and infradian rhythms have a period shorter and longer than 24 hours, respectively. Disruptions in the circadian rhythms contribute to the pathology of cardiovascular diseases, cancer, metabolic syndromes and aging. A transcription/translation feedback loop (TTFL) forms the core of the molecular circadian clock mechanism. Transcription factors, CLOCK or NPAS2 and BMAL1 or BMAL2, form the positive limb of the feedback loop, act in the form of a heterodimer and activate the transcription of core clock genes and clock-controlled genes (involved in key metabolic processes), harboring E-box elements (5'-CACGTG-3') within their promoters. The core clock genes: PER1/2/3 and CRY1/2 which are transcriptional repressors form the negative limb of the feedback loop and interact with the CLOCK|NPAS2-BMAL1|BMAL2 heterodimer inhibiting its activity and thereby negatively regulating their own expression. This heterodimer also activates nuclear receptors NR1D1/2 and RORA/B/G, which form a second feedback loop and which activate and repress BMAL1 transcription, respectively. CRY1 and CRY2 have redundant functions but also differential and selective contributions at least in defining the pace of the SCN circadian clock and its circadian transcriptional outputs. Less potent transcriptional repressor in cerebellum and liver than CRY1, though less effective in lengthening the period of the SCN oscillator. Seems to play a critical role in tuning SCN circadian period by opposing the action of CRY1. With CRY1, dispensable for circadian rhythm generation but necessary for the development of intercellular networks for rhythm synchrony. May mediate circadian regulation of cAMP signaling and gluconeogenesis by blocking glucagon-mediated increases in intracellular cAMP concentrations and in CREB1 phosphorylation. Besides its role in the maintenance of the circadian clock, is also involved in the regulation of other processes. Plays a key role in glucose and lipid metabolism modulation, in part, through the transcriptional regulation of genes involved in these pathways, such as LEP or ACSL4. Represses glucocorticoid receptor NR3C1/GR-induced transcriptional activity by binding to glucocorticoid response elements (GREs). Represses the CLOCK-BMAL1 induced transcription of BHLHE40/DEC1. Represses the CLOCK-BMAL1 induced transcription of NAMPT (By similarity). Represses PPARD and its target genes in the skeletal muscle and limits exercise capacity (By similarity). 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Netherlands)","url":"https://pubmed.ncbi.nlm.nih.gov/28649558","citation_count":4,"is_preprint":false},{"pmid":"37724597","id":"PMC_37724597","title":"Transcription Repression of CRY2 via PER2 Interaction Promotes Adipogenesis.","date":"2023","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/37724597","citation_count":3,"is_preprint":false},{"pmid":"38414323","id":"PMC_38414323","title":"Comparative analysis of locomotor behavior and head diurnal transcriptome regulation by PERIOD and CRY2 in the diamondback moth.","date":"2024","source":"Insect science","url":"https://pubmed.ncbi.nlm.nih.gov/38414323","citation_count":3,"is_preprint":false},{"pmid":"39879691","id":"PMC_39879691","title":"Circadian rhythm gene cryptochrome 2 (Cry2) interacts with lipid metabolism to promote vascular aging.","date":"2025","source":"Archives of gerontology and geriatrics","url":"https://pubmed.ncbi.nlm.nih.gov/39879691","citation_count":3,"is_preprint":false},{"pmid":"27805417","id":"PMC_27805417","title":"Prevalence of cry2-type genes in Bacillus thuringiensis isolates recovered from diverse habitats in India and isolation of a novel cry2Af2 gene toxic to Helicoverpa armigera (cotton boll worm).","date":"2016","source":"Canadian journal of microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/27805417","citation_count":3,"is_preprint":false},{"pmid":"35016737","id":"PMC_35016737","title":"Assessing fitness costs of the resistance of Spodoptera frugiperda (Lepidoptera: Noctuidae) to pyramided Cry1 and Cry2 insecticidal proteins on different host plants.","date":"2022","source":"Bulletin of entomological research","url":"https://pubmed.ncbi.nlm.nih.gov/35016737","citation_count":3,"is_preprint":false},{"pmid":"37492180","id":"PMC_37492180","title":"Expression of CRY2 Gene in the Brain Is Related to Human 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journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/39796036","citation_count":2,"is_preprint":false},{"pmid":"39345648","id":"PMC_39345648","title":"Advancing Clinical Response Against Glioblastoma: Evaluating SHP1705 CRY2 Activator Efficacy in Preclinical Models and Safety in Phase I Trials.","date":"2024","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/39345648","citation_count":2,"is_preprint":false},{"pmid":"19456395","id":"PMC_19456395","title":"Expression of Cry2 in the chicken pineal gland: effects of changes in light-dark conditions.","date":"2009","source":"Annals of the New York Academy of Sciences","url":"https://pubmed.ncbi.nlm.nih.gov/19456395","citation_count":2,"is_preprint":false},{"pmid":"36993226","id":"PMC_36993226","title":"Transcription repression of Cry2 via Per2 interaction promotes adipogenesis.","date":"2023","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/36993226","citation_count":1,"is_preprint":false},{"pmid":"36046448","id":"PMC_36046448","title":"Meta-Analysis of Mechanism of Influence of CRY2 on the Differentiation of Mouse Osteoblast through the Regulation of Wnt/Β-Catenin Signaling Pathway.","date":"2022","source":"BioMed research international","url":"https://pubmed.ncbi.nlm.nih.gov/36046448","citation_count":1,"is_preprint":false},{"pmid":"41490462","id":"PMC_41490462","title":"Cry2 prompts clock oscillation and temporomandibular joint homeostasis under mechanical loading.","date":"2026","source":"Bone & joint research","url":"https://pubmed.ncbi.nlm.nih.gov/41490462","citation_count":1,"is_preprint":false},{"pmid":"41354650","id":"PMC_41354650","title":"Steroid hormone receptors through Cry2 are key players in the circadian clock response to serum.","date":"2025","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/41354650","citation_count":0,"is_preprint":false},{"pmid":"40254427","id":"PMC_40254427","title":"Cry2 Alleviates Cisplatin-Induced Cytotoxicity in Mouse Renal Cortex Tubular Cell Lines.","date":"2025","source":"Biological & pharmaceutical bulletin","url":"https://pubmed.ncbi.nlm.nih.gov/40254427","citation_count":0,"is_preprint":false},{"pmid":"33629313","id":"PMC_33629313","title":"SPRR2C, DEFB4A, WIF1, CRY2, and KRT19 are correlated with the development of atopic eczema.","date":"2021","source":"European review for medical and pharmacological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/33629313","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":49102,"output_tokens":5801,"usd":0.117161,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":14244,"output_tokens":5435,"usd":0.103548,"stage2_stop_reason":"end_turn"},"total_usd":0.220709,"stage1_batch_id":"msgbatch_01ACAxJQabDsht39JXeWG22d","stage2_batch_id":"msgbatch_01QEU22A9U5sbPTgsMzTmvSf","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1999,\n      \"finding\": \"Mice lacking both CRY1 and CRY2 show instantaneous and complete loss of free-running circadian rhythmicity, demonstrating that CRY1 and CRY2 are essential for maintenance of circadian rhythms (not merely photoreceptors). CRY1-null mice show accelerated and CRY2-null mice show delayed free-running locomotor activity periodicity.\",\n      \"method\": \"Genetic knockout (Cry1-/-, Cry2-/-, Cry1-/-/Cry2-/- mice) with locomotor activity monitoring\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean knockout with defined behavioral phenotype, replicated across multiple genotypes in a landmark study\",\n      \"pmids\": [\"10217146\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"CRY2 (and CRY1, PER2) activates transcription of the mouse Bmal1 gene, while BMAL1-CLOCK dimers repress it, establishing an interlocked feedback loop within the circadian clockwork.\",\n      \"method\": \"Transcriptional reporter assays in cell culture; promoter characterization of mBmal1\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — cell-based reporter assays, single lab, but consistent with broader circadian clock model\",\n      \"pmids\": [\"11798163\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"An intact flavin binding domain is required for CRY2 function in suppressing CLOCK/BMAL1-mediated transcription. Mutation of any of the three conserved tryptophan residues in the putative electron transport chain inhibits xCRY2b function, indicating CRY2 requires electron transport through the conserved tryptophan pathway, unlike CRY1 which depends only on the last tryptophan.\",\n      \"method\": \"Site-directed mutagenesis of conserved tryptophan residues; transcriptional suppression assays in Xenopus CRY2\",\n      \"journal\": \"Current biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — mutagenesis with functional readout, but single lab using Xenopus ortholog, not mammalian CRY2\",\n      \"pmids\": [\"11747820\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"CRY2 undergoes proteasomal degradation initiated by a dual-kinase mechanism: DYRK1A phosphorylates Ser557 as a priming event, enabling subsequent GSK-3β-mediated phosphorylation of Ser553, which leads to proteasomal degradation of CRY2. Knockdown of Dyrk1a causes abnormal cytosolic CRY2 accumulation and shortens circadian period.\",\n      \"method\": \"In vitro kinase assays, site-directed mutagenesis (S557A/S553A), RNAi knockdown in cells, circadian period measurement\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro kinase assay, mutagenesis, and cellular phenotype from single lab with multiple orthogonal methods\",\n      \"pmids\": [\"20123978\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Two residues in the C-terminal domain of mouse CRY2, Arg-501 and Lys-503 (within residues 493–512), are essential for direct physical interaction with PER2. Mutation of both residues abolishes CRY2-PER2 interaction.\",\n      \"method\": \"Mammalian two-hybrid assay, co-immunoprecipitation, oligonucleotide-based degenerate PCR mutagenesis\",\n      \"journal\": \"BMC molecular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP and two-hybrid with mutagenesis, single lab\",\n      \"pmids\": [\"20840750\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"CRY1 and CRY2 are both transcriptional repressors within the SCN clockwork, but CRY1 is significantly more potent than CRY2. CRY1 prolongs the interval of transcriptional suppression while CRY2 does not, and CRY2 attenuates the period-lengthening effects of CRY1. Both CRYs dose-dependently lengthen the intrinsic high-frequency SCN rhythm.\",\n      \"method\": \"Genetic epistasis using Fbxl3(Afh) allele in Cry1- and Cry2-deficient mouse backgrounds; SCN bioluminescence recording; wheel-running analysis\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — epistasis analysis across multiple genotypes with two independent phenotypic readouts (behavior and bioluminescence)\",\n      \"pmids\": [\"23616524\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Random mutagenesis identified CRY2-specific residues required for repression of CLOCK-BMAL1. CRY2(G354D) is deficient in clock protein binding required for repression by both CRYs, while CRY2(G351D) shows normal binding but a CRY2-specific repression defect. Overexpression of CRY2(G351D) abolishes circadian rhythmicity, implicating a CRY2-unique repression mechanism.\",\n      \"method\": \"Random mutagenesis screen, cell-based CLOCK-BMAL1 reporter assay, co-immunoprecipitation, circadian rhythm analysis in NIH 3T3 cells\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mutagenesis screen with functional readout and binding assays, single lab\",\n      \"pmids\": [\"19687303\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"FBXW7 is an E3 ubiquitin ligase that targets CRY2 for proteasomal degradation by binding directly to phosphorylated Thr300 of CRY2, enhancing CRY2 ubiquitination and accelerating CRY2 turnover.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assays, site-directed mutagenesis (Thr300), cycloheximide chase; biochemical and cell biology analyses\",\n      \"journal\": \"Molecular cancer therapeutics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct binding and ubiquitination assays with phospho-site mutagenesis, single lab\",\n      \"pmids\": [\"25855785\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"CRY2 functions as a component of an FBXL3-containing SCF E3 ubiquitin ligase complex that recruits T58-phosphorylated c-MYC for ubiquitylation and degradation. CRY1 cannot substitute for CRY2 in this function. This represents a circadian mechanism for control of cell proliferation via c-MYC turnover.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitylation assays, phospho-specific recruitment assays; genetic complementation showing CRY1 cannot substitute\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, in-cell ubiquitination assays, isoform specificity established, multiple orthogonal approaches\",\n      \"pmids\": [\"27840026\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"CRY1 and CRY2 broadly interact with nuclear hormone receptors and serve as corepressors for many NRs, binding independently of other core clock factors to genomic sites enriched for NR recognition motifs, contributing to circadian regulation of drug metabolism.\",\n      \"method\": \"Genomic binding analysis (ChIP-related), co-repressor functional assays; interaction studies with multiple NRs\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genomic binding and functional corepressor assays, single lab with multiple NRs tested\",\n      \"pmids\": [\"28751364\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"CRY2, but not CRY1, specifically interacts with Bclaf1 to stabilize mRNAs encoding cyclin D1 and Tmem176b, regulating circadian patterns of myoblast proliferation and myogenic cell fusion. Cry2-/- myoblasts show premature cell cycle exit and form short myotubes, and muscle regeneration is impaired in Cry2-/- mice.\",\n      \"method\": \"Co-immunoprecipitation of CRY2-Bclaf1 complex, Cry2-/- mouse model, mRNA stability assays, single myofiber analysis, muscle regeneration assays\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, knockout mouse with defined phenotype, mRNA stability assays, Bclaf1 knockdown phenocopy, multiple orthogonal methods\",\n      \"pmids\": [\"29466738\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Two cancer-associated CRY2 missense mutations (D325H and S510L in mouse) suppress P53 target-gene expression and accelerate growth of primary fibroblasts expressing high c-MYC. The mutations have divergent impacts on circadian rhythms and on CRY2's ability to interact with SCF-FBXL3, and neither affects steady-state levels of overexpressed c-MYC.\",\n      \"method\": \"Stable expression of CRY2 mutants in primary mouse fibroblasts, P53 target gene expression analysis, circadian rhythm assays, co-immunoprecipitation with FBXL3\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — cell-based functional assays with defined mutants, Co-IP, single lab with multiple readouts\",\n      \"pmids\": [\"34183418\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"X-ray crystal structure of mammalian CRY2 in complex with the small molecule SHP656 reveals that compound binding is compatible with the intrinsic CRY2 gatekeeper W417 'in' orientation. The gatekeeper residue W417 and lid loop interactions are important for CRY2 isoform selectivity of this compound. SHP656 lengthened cellular circadian period in a CRY2-dependent manner.\",\n      \"method\": \"X-ray crystallography, molecular dynamics simulations, circadian period assays in CRY2-dependent manner, mutagenesis of W417\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure with mutagenesis validation and cellular functional assay, single lab\",\n      \"pmids\": [\"36161947\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Human CRY2 knockout cells generated by CRISPR/Cas9 show long-period circadian rhythms, while CRY1/CRY2 double knockout cells are arrhythmic, consistent with mouse knockout models and confirming CRY2's period-setting role in human cells.\",\n      \"method\": \"CRISPR/Cas9 knockout of CRY2 in human U-2 OS cells; circadian bioluminescence recording\",\n      \"journal\": \"Frontiers in physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean CRISPR KO with defined circadian phenotype, single lab, consistent with prior mouse data\",\n      \"pmids\": [\"31143130\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"CRY2 variant p.Ser420Phe has reduced affinity for PER2 and defective nuclear translocation, resulting in reduced repression of CLOCK:BMAL1-driven transcription. This variant is unexpectedly resistant to degradation via FBXL3 and FBXL21 (canonical proteasomal pathway), indicating Ser-420 is required for E3 ligase interaction. Wild-type CRY2 (and this variant) can also be degraded via a lysosomal pathway. The variant causes a ~7 h shorter circadian period in Cry1-/-Cry2-/- MEFs.\",\n      \"method\": \"Site-directed mutagenesis, co-immunoprecipitation with PER2/FBXL3/FBXL21, nuclear localization imaging, proteasomal/lysosomal inhibitor experiments, complementation in Cry1-/-Cry2-/- double KO MEFs\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — mutagenesis, multiple Co-IP experiments, degradation pathway dissection with inhibitors, functional complementation, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"37951306\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"CRY2 represses CLOCK/BMAL1-induced transcription via a critical Cys432 residue that mediates PER2 heterodimer formation. The C432 mutation disrupts PER2 association without affecting BMAL1 binding. This CRY2-PER2 repression complex is required for transcriptional repression of Wnt pathway components to promote adipogenesis.\",\n      \"method\": \"Site-directed mutagenesis (C432), co-immunoprecipitation, transcriptional reporter assays, adipogenic differentiation assays, CRY2 KD/OE in preadipocytes, KL001 stabilization\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — mutagenesis identifying specific residue, Co-IP, functional adipogenesis assay, multiple orthogonal methods in single lab\",\n      \"pmids\": [\"37724597\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"FBXW7 promotes CRY2 ubiquitin-mediated degradation and thereby promotes trophoblast migration and invasion. The lncRNA MALAT1 recruits FBXW7 to impair CRY2 protein stability; reduced MALAT1 leads to CRY2 accumulation and suppressed migration/invasion.\",\n      \"method\": \"RNA pull-down, co-immunoprecipitation of FBXW7-CRY2, CRY2 protein stability assays, trophoblast migration/invasion assays\",\n      \"journal\": \"Journal of cellular physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — Co-IP and functional assays, single lab, consistent with FBXW7-CRY2 interaction from prior work\",\n      \"pmids\": [\"32776544\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"CRY2 overexpression impairs trophoblast migration and invasion by inhibiting the c-Myc-BMAL1 pathway. c-Myc binds the BMAL1 promoter to induce BMAL1 transcription, which activates MMP2/9. CRY2 suppresses this c-Myc-BMAL1-MMP2/9 axis.\",\n      \"method\": \"Luciferase reporter assay, chromatin immunoprecipitation (c-Myc on BMAL1 promoter), wound healing and Transwell invasion assays, CRY2 overexpression/knockdown in HTR-8/SVneo cells\",\n      \"journal\": \"Journal of biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP, reporter assay, and functional migration assays with CRY2 OE/KD, single lab\",\n      \"pmids\": [\"31536114\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Loss of CRY2 in satellite cells enhances muscle regeneration by activating ERK1/2 signaling and ETS1, which binds the PAX7 promoter to induce PAX7 transcription and increase satellite cell proliferation.\",\n      \"method\": \"Satellite cell/skeletal muscle lineage-specific Cry2 knockout mice (CRY2scko), immunostaining, single myofiber analysis, ERK1/2 activation assays, ChIP (ETS1 on PAX7 promoter)\",\n      \"journal\": \"MedComm\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — tissue-specific KO mouse, ChIP, signaling pathway characterization, single lab\",\n      \"pmids\": [\"36636367\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"CRY2 physically interacts with TRPC1, and this interaction is detected by co-immunoprecipitation. Both proteins co-translocate to the nucleus following pulsed electromagnetic field (PEMF) exposure. CRY2 overexpression enhances PEMF-induced myogenic responses, while CRY2 silencing reduces them. Reducing FAD content by silencing riboflavin kinase attenuates PEMF responsiveness.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence co-localization, CRY2 overexpression/knockdown, riboflavin kinase silencing, PEMF exposure assays in myoblasts\",\n      \"journal\": \"Cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — Co-IP and co-localization with functional OE/KD assays, single lab, novel interaction\",\n      \"pmids\": [\"39937022\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"CRY2 plays a central role in serum-induced circadian clock phase resetting. Steroid hormone receptors (including sex-hormone receptors) are key mediators of serum-induced phase resetting, and CRY2 mediates their effect on clock phase independently of its role in period-length determination.\",\n      \"method\": \"Circa-SCOPE high-throughput single-cell phase transition curve analysis; CRY2 perturbation experiments; steroid hormone receptor pathway analysis\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — novel high-throughput single-cell method with CRY2 perturbation, single lab\",\n      \"pmids\": [\"41354650\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"CRY2 overexpression in mouse renal tubular cells alleviates cisplatin-induced cytotoxicity by reducing platinum accumulation through upregulation of efflux transporters ATP7A and MRP2.\",\n      \"method\": \"CRY2 overexpression in MuRTE61 cells, cell viability assays, platinum content measurement, western blot of ATP7A and MRP2\",\n      \"journal\": \"Biological & pharmaceutical bulletin\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single OE experiment with defined readout, single lab, single method for mechanism\",\n      \"pmids\": [\"40254427\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"CRY2 nuclear-cytoplasmic shuttling in chondrocytes depends on ROCK activation and actin polymerization under mechanical loading. Inhibition of ROCK causes actin depolymerization and partially blocks CRY2 nuclear-cytoplasmic trafficking. Knockdown of CRY2 attenuates mechanical loading-sustained circadian oscillations and cartilage homeostasis.\",\n      \"method\": \"ROCK inhibitor treatment, actin polymerization assays, CRY2 localization imaging (nuclear-cytoplasmic shuttling), CRY2 knockdown in chondrocytes and condyle explants\",\n      \"journal\": \"Bone & joint research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — direct localization with mechanistic inhibitor experiments and functional KD readout, single lab\",\n      \"pmids\": [\"41490462\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"Mammalian CRY2 is a core circadian transcriptional repressor that suppresses CLOCK/BMAL1-driven transcription via a PER2-interacting C-terminal domain (requiring residues including Arg-501, Lys-503, and Cys-432), undergoes circadian-regulated proteasomal degradation initiated by DYRK1A-mediated priming phosphorylation at Ser557 followed by GSK-3β phosphorylation at Ser553 (with FBXL3 and FBXL21 as E3 ligases and a secondary lysosomal degradation pathway), and performs non-redundant clock-output functions including acting as an adaptor within SCF-FBXL3 to recruit phospho-T58 c-MYC for ubiquitylation, interacting with Bclaf1 to stabilize myogenic mRNAs, serving as a corepressor for nuclear hormone receptors, and mediating serum-induced phase resetting via steroid hormone receptors.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CRY2 is a core component of the mammalian circadian clockwork that acts as a transcriptional repressor of the CLOCK/BMAL1 complex, and its loss alters or, in combination with CRY1, abolishes free-running circadian rhythmicity [#0, #13]. Repression depends on a C-terminal domain that mediates direct heterodimerization with PER2 through residues including Arg-501/Lys-503 and Cys-432, the latter disrupting PER2 association without affecting BMAL1 binding, and on additional residues such as Gly-351 that define a CRY2-specific repression activity distinct from CRY1 [#4, #15, #6]; an intact flavin/electron-transport pathway is also required for CLOCK/BMAL1 suppression [#2]. CRY2 abundance is set by regulated proteolysis: DYRK1A primes Ser557 phosphorylation to enable GSK-3\\u03b2 phosphorylation of Ser553 and proteasomal turnover, while FBXL3/FBXL21 and FBXW7 (the latter binding phospho-Thr300) target CRY2 for ubiquitin-mediated degradation, with Ser-420 required for E3 ligase engagement and a parallel lysosomal route also operating [#3, #7, #14]. Beyond the canonical loop, CRY2 performs non-redundant, isoform-specific output functions that CRY1 cannot substitute: it serves as an adaptor in an SCF-FBXL3 complex to recruit phospho-T58 c-MYC for ubiquitylation, links the clock to cell proliferation and P53 target regulation, and binds Bclaf1 to stabilize myogenic mRNAs controlling myoblast proliferation and muscle regeneration [#8, #11, #10]. CRY2 additionally acts as a corepressor for nuclear hormone receptors at NR-motif-enriched genomic sites and mediates steroid-hormone-receptor-dependent serum-induced phase resetting independently of its period-setting role [#9, #20]. A crystal structure of CRY2 bound to the period-lengthening compound SHP656 defines a gatekeeper W417 conformation underlying isoform-selective small-molecule modulation [#12].\",\n  \"teleology\": [\n    {\n      \"year\": 1999,\n      \"claim\": \"Established that CRY2 is not merely a photoreceptor but an essential clock component, resolving whether cryptochromes maintain mammalian rhythmicity.\",\n      \"evidence\": \"Single and double Cry1/Cry2 knockout mice with locomotor activity monitoring\",\n      \"pmids\": [\"10217146\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not define the molecular mechanism of repression\", \"Behavioral phenotype does not localize CRY2 action to specific molecular partners\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Addressed whether the flavin/electron-transport apparatus is functionally required for transcriptional repression, distinguishing CRY2 from CRY1.\",\n      \"evidence\": \"Site-directed mutagenesis of conserved tryptophans with transcriptional suppression assays in Xenopus CRY2\",\n      \"pmids\": [\"11747820\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Used Xenopus ortholog, not mammalian CRY2\", \"Did not establish a photochemical or redox mechanism in mammalian cells\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Defined the interlocked feedback architecture by showing CRY2 activates Bmal1 transcription while CLOCK/BMAL1 represses it.\",\n      \"evidence\": \"Transcriptional reporter assays and mBmal1 promoter characterization in cell culture\",\n      \"pmids\": [\"11798163\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Reporter-based; in vivo contribution to the loop not directly tested\", \"Mechanism of CRY2-mediated activation unresolved\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Distinguished clock-protein binding from a CRY2-unique repression step, showing CRY2 has a repression mechanism separable from complex assembly.\",\n      \"evidence\": \"Random mutagenesis screen (G354D, G351D), reporter assays, Co-IP and rhythm analysis in NIH 3T3 cells\",\n      \"pmids\": [\"19687303\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular nature of the CRY2-specific repression defect not defined\", \"Single lab, cell-based\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Mapped the PER2 interaction surface and the priming-phosphorylation degradation switch, defining how CRY2 abundance is timed.\",\n      \"evidence\": \"Two-hybrid/Co-IP mutagenesis (R501/K503) and in vitro DYRK1A/GSK-3\\u03b2 kinase assays with S557A/S553A mutants and RNAi\",\n      \"pmids\": [\"20840750\", \"20123978\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not identify the cognate E3 ligase acting on phospho-CRY2\", \"Kinase priming hierarchy shown in vitro and cells but not in vivo timing\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Quantified the functional asymmetry between CRY1 and CRY2, showing CRY2 is a weaker repressor that modulates CRY1 action.\",\n      \"evidence\": \"Fbxl3(Afh) epistasis across Cry-deficient backgrounds with SCN bioluminescence and wheel-running\",\n      \"pmids\": [\"23616524\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of differential repressor potency not resolved\", \"Does not explain CRY2's non-redundant output roles\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Identified FBXW7 as a phospho-Thr300-dependent E3 ligase for CRY2, expanding the degradation machinery beyond FBXL3/FBXL21.\",\n      \"evidence\": \"Co-IP, ubiquitination assays, Thr300 mutagenesis and cycloheximide chase\",\n      \"pmids\": [\"25855785\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Kinase phosphorylating Thr300 not identified\", \"Relative contribution versus FBXL3/FBXL21 unresolved\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Revealed a non-redundant output function: CRY2 acts as an SCF-FBXL3 adaptor recruiting phospho-T58 c-MYC, linking the clock to proliferation.\",\n      \"evidence\": \"Co-IP, in-cell ubiquitylation, phospho-specific recruitment and CRY1 non-complementation\",\n      \"pmids\": [\"27840026\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo tumor-suppressive role not established here\", \"Structural basis of the c-MYC recruitment specificity unknown\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Showed CRY2 broadly corepresses nuclear hormone receptors independently of core clock factors, connecting the clock to drug metabolism.\",\n      \"evidence\": \"Genomic binding analysis and corepressor functional assays across multiple NRs\",\n      \"pmids\": [\"28751364\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"CRY2-versus-CRY1 specificity at NR sites not dissected\", \"Direct versus indirect genomic recruitment unclear\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Demonstrated a post-transcriptional output role: CRY2-specific binding to Bclaf1 stabilizes myogenic mRNAs and controls muscle regeneration.\",\n      \"evidence\": \"Co-IP, Cry2-/- mice, mRNA stability assays, single myofiber and regeneration assays\",\n      \"pmids\": [\"29466738\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism of CRY2/Bclaf1 mRNA selectivity unknown\", \"Relationship to CRY2's nuclear repressor role not integrated\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Confirmed CRY2's period-setting role in human cells, validating cross-species conservation of the phenotype.\",\n      \"evidence\": \"CRISPR/Cas9 CRY2 and CRY1/CRY2 knockouts in U-2 OS cells with bioluminescence\",\n      \"pmids\": [\"31143130\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No molecular dissection beyond period phenotype\", \"Output functions not tested in this system\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Connected CRY2 turnover to trophoblast behavior, showing FBXW7-driven degradation and CRY2 suppression of the c-Myc-BMAL1-MMP axis.\",\n      \"evidence\": \"RNA pull-down, Co-IP, stability and migration/invasion assays; ChIP of c-Myc on BMAL1 promoter in HTR-8/SVneo cells\",\n      \"pmids\": [\"32776544\", \"31536114\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"In vivo relevance to placentation not established\", \"Single cell-line systems\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Linked cancer-associated CRY2 missense variants to P53 target suppression and c-MYC-dependent growth, implicating CRY2 in tumor biology.\",\n      \"evidence\": \"Stable CRY2 mutant expression in primary fibroblasts, P53 target and circadian assays, FBXL3 Co-IP\",\n      \"pmids\": [\"34183418\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism connecting variants to P53 targets undefined\", \"No in vivo tumorigenesis data\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Provided a structural basis for isoform-selective small-molecule modulation, defining the W417 gatekeeper conformation.\",\n      \"evidence\": \"X-ray crystallography of CRY2-SHP656, MD simulations, W417 mutagenesis and CRY2-dependent period assays\",\n      \"pmids\": [\"36161947\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structure of full repressor complex with PER2/CLOCK/BMAL1 not resolved\", \"Endogenous ligand state not addressed\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Resolved the Cys432/Ser420 determinants of PER2 binding, repression and E3 engagement, and extended CRY2 repression to Wnt-driven adipogenesis and ERK/ETS1-PAX7 muscle programs.\",\n      \"evidence\": \"C432 and S420F mutagenesis with PER2/FBXL3/FBXL21 Co-IP, localization imaging, degradation inhibitors, adipogenesis assays, and satellite-cell Cry2 KO with ChIP\",\n      \"pmids\": [\"37724597\", \"37951306\", \"36636367\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How a single residue couples binding, localization and degradation not fully mechanized\", \"Tissue-specific output mechanisms not unified\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identified period-independent and signaling-coupled roles: CRY2 mediates steroid-hormone-receptor-dependent phase resetting, interacts with TRPC1 in electromagnetic-field responses, and modulates renal drug efflux.\",\n      \"evidence\": \"Single-cell phase-transition (Circa-SCOPE) with CRY2 perturbation; Co-IP/co-localization with TRPC1 and riboflavin-kinase silencing in myoblasts; CRY2 overexpression with ATP7A/MRP2 readouts in renal cells\",\n      \"pmids\": [\"41354650\", \"39937022\", \"40254427\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"TRPC1 interaction is single-lab without reciprocal in vivo validation\", \"Renal efflux role rests on a single overexpression experiment (Low confidence)\", \"Molecular link between steroid receptors and CRY2 phase resetting undefined\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Showed CRY2 nuclear-cytoplasmic shuttling is mechanosensitive, coupling ROCK/actin dynamics to clock oscillations in cartilage.\",\n      \"evidence\": \"ROCK inhibition, actin polymerization assays, CRY2 localization imaging and knockdown in chondrocytes and condyle explants\",\n      \"pmids\": [\"41490462\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct molecular link between actin and CRY2 transport unknown\", \"In vivo cartilage relevance from explant model only\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How CRY2's distinct activities — transcriptional repression, E3-adaptor function, mRNA stabilization, NR corepression, and mechano-/hormone-coupled localization — are integrated and isoform-specifically partitioned from CRY1 remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structure of the assembled CRY2-PER2-CLOCK/BMAL1 repressor complex\", \"Mechanism determining when CRY2 acts as repressor versus SCF adaptor versus RNA-stabilizing factor unknown\", \"Endogenous flavin/redox cofactor function in mammalian CRY2 not established\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [0, 1, 6, 9, 15]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [8, 11]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [8, 10]},\n      {\"term_id\": \"GO:0003723\", \"supporting_discovery_ids\": [10]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [9, 15]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [14, 22]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [3, 22]},\n      {\"term_id\": \"GO:0005654\", \"supporting_discovery_ids\": [9, 15]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-9909396\", \"supporting_discovery_ids\": [0, 5, 13, 20]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [1, 6, 9, 15]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [3, 7, 8, 14]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [8, 10, 11]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [10, 15, 18]}\n    ],\n    \"complexes\": [\n      \"CRY2-PER2 repressor complex\",\n      \"SCF-FBXL3 E3 ubiquitin ligase complex\"\n    ],\n    \"partners\": [\n      \"PER2\",\n      \"BMAL1\",\n      \"CLOCK\",\n      \"FBXL3\",\n      \"FBXW7\",\n      \"c-MYC\",\n      \"Bclaf1\",\n      \"TRPC1\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}