{"gene":"NUDT16L1","run_date":"2026-06-10T05:19:52","timeline":{"discoveries":[{"year":2017,"finding":"TIRR (NUDT16L1) directly binds the tandem Tudor domain of 53BP1 and masks its H4K20me2 histone methyl-lysine binding motif, preventing 53BP1 recruitment to DNA double-strand breaks. Upon DNA damage, ATM phosphorylates 53BP1 and recruits RIF1 to dissociate the 53BP1-TIRR complex. Overexpression of TIRR impedes 53BP1 localization to DSBs, while depletion of TIRR destabilizes 53BP1 in the nuclear-soluble fraction.","method":"Co-immunoprecipitation, overexpression/depletion functional assays, live-cell imaging of DSB foci","journal":"Nature","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, multiple orthogonal functional assays, replicated by multiple subsequent independent labs","pmids":["28241136"],"is_preprint":false},{"year":2018,"finding":"Crystal structure (1.76 Å) of TIRR in complex with the 53BP1 tandem Tudor domain reveals that the N-terminal region (residues 10–24) and the L8-loop of TIRR interact with 53BP1 Tudor through three loops (L1, L3, L1'), blocking the H4K20me2-binding surface. A TIRR-specific histidine (H106), absent from the homolog NUDT16, is essential for 53BP1 Tudor binding; mutations mimicking TIRR binding modules restore disrupted NUDT16-53BP1 Tudor interaction.","method":"X-ray crystallography, site-directed mutagenesis, binding assays","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1 / Strong — high-resolution crystal structure with mutagenesis validation, independently corroborated by two contemporaneous structural studies","pmids":["29844495"],"is_preprint":false},{"year":2018,"finding":"X-ray crystal structure of TIRR bound to 53BP1 tandem Tudor domain reveals that an essential TIRR arginine residue is central to an intricate binding area that blocks the methylated-chromatin-binding surface of 53BP1. A 53BP1 separation-of-function mutation abolishing TIRR-mediated regulation renders 53BP1 hyperactive at DSBs. TIRR-interacting RNA molecules relieve this inhibition, providing proof-of-principle of RNA-triggered 53BP1 recruitment to DSBs.","method":"X-ray crystallography, site-directed mutagenesis, cell-based DSB foci assays, RNA binding assays","journal":"Nature structural & molecular biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure with mutagenesis and functional cell-based validation in one study, corroborated by independent structural studies","pmids":["29967538"],"is_preprint":false},{"year":2018,"finding":"Crystal structure of TIRR–53BP1 tandem Tudor domain complex shows three TIRR loops masking the methylated lysine-binding pocket of 53BP1 TTD, competing with histone H4K20 methylation. Key interaction residues were mapped and their mutation abolishes complex formation. NUDT16 does not directly interact with 53BP1 due to absence of key binding residues. TIRR suppresses relocation of 53BP1 to DNA lesions and 53BP1-dependent DNA damage repair.","method":"X-ray crystallography, site-directed mutagenesis, Co-IP, DSB repair assays","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure with mutagenesis and functional validation; explicitly demonstrates NUDT16 lacks key binding residues as a negative control","pmids":["30002377"],"is_preprint":false},{"year":2021,"finding":"TIRR inhibits complex formation between the Tudor domain of 53BP1 and dimethylated p53 (K382me2), thereby suppressing p53 transcriptional activation of target genes. Loss of TIRR causes an aberrant increase in p53 gene transactivation, affecting p53-mediated cell-fate programs. TIRR depletion is selectively not tolerated in p53-proficient tumors.","method":"Biochemical binding assays (Tudor domain–K382me2 p53 interaction), gene expression analysis upon TIRR loss, cell viability assays","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 / Moderate — biochemical complex reconstitution with cellular functional readout, single lab with multiple orthogonal methods","pmids":["33961797"],"is_preprint":false},{"year":2022,"finding":"RNA with a hairpin secondary structure transcribed at DSBs by RNA polymerase II promotes TIRR/53BP1 complex dissociation. This hairpin RNA binds to the same residues on TIRR as 53BP1, providing the mechanistic basis for RNA-driven complex separation after DNA damage.","method":"RNA binding assays, mutagenesis mapping of RNA-TIRR interaction surface, cell-based TIRR/53BP1 dissociation assays, RNA polymerase II inhibition","journal":"Cell reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct RNA–protein binding mapped to specific residues with functional complex dissociation assay, single lab","pmids":["36288694"],"is_preprint":false},{"year":2024,"finding":"NEAT1 long non-coding RNA (specifically the short isoform NEAT1_1, enriched in G1 phase) is the primary RNA partner of TIRR within cells, identified by iCLIP. NEAT1_1 binding destabilizes the TIRR/53BP1 complex, promoting 53BP1 function in a cell-cycle-dependent manner. TDP-43 modulates the TIRR/53BP1 complex by promoting NEAT1_1 production.","method":"iCLIP (individual-nucleotide resolution UV crosslinking and immunoprecipitation), Co-IP, cell-cycle synchronization, TDP-43 functional assays","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — iCLIP identifies direct RNA–protein contacts; functional disruption of TIRR/53BP1 complex validated, single lab","pmids":["39349456"],"is_preprint":false},{"year":2024,"finding":"DTX3L ubiquitinates TIRR at lysine 187, facilitating XPO1-mediated nuclear export and subsequent degradation of TIRR upon DNA damage. This relieves TIRR-mediated inhibition of 53BP1, regulating NHEJ pathway activity and PARP inhibitor sensitivity. DTX3L overexpression (as in prostate cancers) decreases TIRR levels, impairs HR, and induces chromosomal instability.","method":"Ubiquitination assays, nuclear export inhibition (XPO1 inhibitor), site-directed mutagenesis (K187), co-immunoprecipitation, cellular fractionation, PARP inhibitor sensitivity assays","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — specific ubiquitination site identified with mutagenesis, export mechanism validated with inhibitor, multiple orthogonal functional readouts","pmids":["39632881"],"is_preprint":false},{"year":2024,"finding":"TIRR selectively binds a subset of mRNAs in response to DNA damage and interacts with the nuclear export protein Exportin-1 (XPO1/CRM1) through a nuclear export signal. TIRR and TIRR-bound RNA co-localize with processing bodies (P-bodies); TIRR depletion results in nuclear RNA retention and impaired P-body formation, linking TIRR's RNA-binding activity to mRNA nuclear export and storage during the DDR.","method":"RNA immunoprecipitation, co-immunoprecipitation with XPO1, fluorescence microscopy (P-body co-localization), TIRR depletion with mRNA export/P-body formation readouts","journal":"Nucleic acids research","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — direct protein–RNA and protein–XPO1 interactions demonstrated, P-body role validated by depletion, single lab single study","pmids":["39119906"],"is_preprint":false},{"year":2024,"finding":"NUDT16L1 localizes to mitochondria in colon cancer cells, where it prevents mitochondrial DNA leakage upon ferroptosis induction. It promotes ferroptosis insensitivity by binding directly to NAD-capped RNAs and indirectly enhancing expression of ferroptosis repressor and mitochondrial genes through MALAT1.","method":"Subcellular fractionation/mitochondrial localization, NAD-capped RNA binding assays, NUDT16L1 overexpression/knockdown with ferroptosis assays, transgenic mouse model","journal":"Redox biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct mitochondrial localization with functional consequence (mtDNA leakage), direct NAD-capped RNA binding demonstrated, in vivo model corroboration","pmids":["39317106"],"is_preprint":false},{"year":2024,"finding":"Deletion of TIRR in mice selectively activates p53, protecting against cancer but causing systemic metabolic imbalance (overweight, insulin resistance). These metabolic and oncoprotective effects are dependent on p53. Tissue-specific models indicate glucose homeostasis is regulated primarily by TIRR expression in adipose tissue, and orexigenesis by TIRR expression in the CNS.","method":"Conditional/tissue-specific mouse knockout models, metabolic phenotyping, p53 deletion epistasis","journal":"Cell reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — tissue-specific KO with epistasis to p53 establishes pathway placement; in vivo mouse models with defined metabolic phenotypes","pmids":["38861384"],"is_preprint":false},{"year":2026,"finding":"EGR1 transcription factor directly binds the promoter of NUDT16L1 and activates its expression, establishing NUDT16L1 as a downstream effector of EGR1. Overexpression of NUDT16L1 alone reduces ROS and prevents cell death in spermatogonial stem cells under oxidative stress; silencing NUDT16L1 abolishes EGR1's protective effects against testicular ischemia-reperfusion injury in mice.","method":"Chromatin immunoprecipitation (ChIP), luciferase reporter assay, NUDT16L1 overexpression/knockdown with ROS/apoptosis readouts, mouse testicular IR injury model","journal":"Cell biology international","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP and luciferase confirm direct transcriptional regulation; epistasis (KD abolishes EGR1 effect; OE mimics it) places NUDT16L1 downstream, single lab","pmids":["41521514"],"is_preprint":false}],"current_model":"NUDT16L1/TIRR is an RNA-binding protein that directly binds the tandem Tudor domain of 53BP1 (via its N-terminal region, L8-loop, and a critical H106 residue), masking its H4K20me2 and K382me2-p53 binding surfaces to inhibit 53BP1 recruitment to DNA double-strand breaks and 53BP1-mediated p53 activation; upon DNA damage, DSB-derived hairpin RNA and NEAT1_1 lncRNA compete for the same TIRR surface to dissociate the TIRR/53BP1 complex, while DTX3L-mediated ubiquitination of TIRR at K187 drives XPO1-dependent nuclear export and proteolytic degradation of TIRR, collectively relieving 53BP1 inhibition; TIRR additionally functions as an mRNA export regulator via XPO1 and modulates P-body formation, and in certain cancer contexts localizes to mitochondria to bind NAD-capped RNAs and suppress ferroptosis, with the EGR1 transcription factor acting as an upstream activator of NUDT16L1 transcription in spermatogonial stem cells."},"narrative":{"mechanistic_narrative":"NUDT16L1 (TIRR) is an RNA-binding regulator of the DNA damage response that controls 53BP1 activity by directly binding its tandem Tudor domain [PMID:28241136]. High-resolution crystal structures show that the TIRR N-terminal region and L8-loop engage 53BP1 Tudor through three loops, masking the methyl-lysine binding surface that 53BP1 otherwise uses to read H4K20me2 chromatin marks; a TIRR-specific histidine (H106) and an essential arginine are central to this interface, and these residues distinguish TIRR from its homolog NUDT16, which does not bind 53BP1 [PMID:29844495, PMID:29967538, PMID:30002377]. By occluding this surface, TIRR prevents 53BP1 recruitment to double-strand breaks and dampens DNA repair, while a parallel interaction blocks 53BP1 binding to dimethylated p53 (K382me2), suppressing p53-dependent transcription; loss of TIRR hyperactivates p53 and is selectively lethal in p53-proficient tumors [PMID:28241136, PMID:33961797]. TIRR inhibition is relieved upon DNA damage through competing RNAs that bind the same TIRR surface as 53BP1—DSB-derived hairpin RNAs and the NEAT1_1 lncRNA isoform dissociate the TIRR/53BP1 complex in a cell-cycle-dependent manner [PMID:29967538, PMID:36288694, PMID:39349456]—and through DTX3L-mediated ubiquitination of TIRR at K187, which drives XPO1-dependent nuclear export and degradation [PMID:39632881]. Beyond 53BP1 regulation, TIRR couples to mRNA metabolism, binding selected mRNAs and the export factor XPO1 to direct nuclear RNA export and P-body formation during the DDR [PMID:39119906]. In mice, TIRR loss activates p53 to confer cancer protection at the cost of systemic metabolic imbalance [PMID:38861384]. In specific cancer contexts NUDT16L1 localizes to mitochondria, binds NAD-capped RNAs, and suppresses ferroptosis [PMID:39317106], and its transcription is directly activated by EGR1 to protect spermatogonial stem cells from oxidative stress [PMID:41521514].","teleology":[{"year":2017,"claim":"Established that TIRR is a direct negative regulator of 53BP1, answering how 53BP1 recruitment to breaks is held in check before damage.","evidence":"Co-IP, overexpression/depletion functional assays, and live-cell DSB foci imaging in human cells","pmids":["28241136"],"confidence":"High","gaps":["Did not define the atomic basis of the TIRR/53BP1 interface","Mechanism of RNA-driven dissociation not yet established"]},{"year":2018,"claim":"Crystal structures resolved how TIRR masks the 53BP1 Tudor methyl-lysine pocket and identified the specific residues (H106, an essential arginine) that confer specificity, explaining why the homolog NUDT16 cannot bind.","evidence":"X-ray crystallography of TIRR-53BP1 Tudor complexes with site-directed mutagenesis, DSB foci and repair assays, and RNA binding assays across three independent studies","pmids":["29844495","29967538","30002377"],"confidence":"High","gaps":["Identity of the physiological dissociating RNA species in cells not yet defined","How damage signaling triggers dissociation in vivo not fully resolved"]},{"year":2021,"claim":"Extended TIRR's regulatory reach beyond chromatin to p53, showing it blocks the 53BP1 Tudor-K382me2-p53 interaction and thereby restrains p53 transactivation, defining a synthetic vulnerability in p53-proficient tumors.","evidence":"Biochemical Tudor-K382me2 binding assays, gene expression analysis on TIRR loss, and cell viability assays","pmids":["33961797"],"confidence":"High","gaps":["Cell-type specificity of the p53 axis not fully mapped","Did not address how this is coordinated with the DSB-recruitment function"]},{"year":2022,"claim":"Identified the molecular trigger for relieving TIRR inhibition after damage: DSB-transcribed hairpin RNAs bind the same TIRR residues as 53BP1 and competitively dissociate the complex.","evidence":"RNA binding assays, mutagenesis mapping of the RNA-TIRR surface, RNA Pol II inhibition, and cell-based dissociation assays","pmids":["36288694"],"confidence":"Medium","gaps":["Single lab; reciprocal validation of the in-cell hairpin species limited","Kinetics relative to RIF1/ATM-driven dissociation not resolved"]},{"year":2024,"claim":"Defined NEAT1_1 lncRNA as the primary cellular RNA partner of TIRR and linked its TDP-43-dependent production to cell-cycle-dependent control of 53BP1.","evidence":"iCLIP, Co-IP, cell-cycle synchronization, and TDP-43 functional assays","pmids":["39349456"],"confidence":"Medium","gaps":["Single lab; relationship between NEAT1_1 and DSB hairpin RNA competition not reconciled","Quantitative contribution of TDP-43 in vivo not established"]},{"year":2024,"claim":"Revealed a degradative arm of TIRR regulation, showing DTX3L ubiquitinates TIRR at K187 to drive XPO1-dependent nuclear export and degradation, coupling TIRR turnover to repair-pathway choice and PARP inhibitor sensitivity.","evidence":"Ubiquitination assays, K187 mutagenesis, XPO1 inhibition, fractionation, and PARP inhibitor sensitivity assays","pmids":["39632881"],"confidence":"High","gaps":["How RNA-driven and ubiquitination-driven dissociation are temporally coordinated unclear","Generality of DTX3L overexpression effect across cancer types not fully tested"]},{"year":2024,"claim":"Expanded TIRR function to mRNA metabolism, demonstrating XPO1-coupled mRNA export and P-body involvement during the DDR.","evidence":"RNA-IP, Co-IP with XPO1, fluorescence microscopy of P-body co-localization, and depletion readouts of RNA export","pmids":["39119906"],"confidence":"Medium","gaps":["Single lab single study","Identity and fate of the exported mRNA cargo not defined"]},{"year":2024,"claim":"Identified a context-specific mitochondrial role for NUDT16L1 in binding NAD-capped RNAs and suppressing ferroptosis in colon cancer.","evidence":"Subcellular fractionation, NAD-capped RNA binding assays, ferroptosis assays, and a transgenic mouse model","pmids":["39317106"],"confidence":"Medium","gaps":["Mechanistic link to the nuclear 53BP1/RNA functions unclear","Generality beyond colon cancer not established"]},{"year":2024,"claim":"Placed TIRR upstream of organismal phenotypes via p53, showing tissue-specific knockouts produce cancer protection alongside metabolic imbalance in a p53-dependent manner.","evidence":"Conditional/tissue-specific mouse knockouts, metabolic phenotyping, and p53 deletion epistasis","pmids":["38861384"],"confidence":"Medium","gaps":["Molecular link between TIRR and tissue metabolism beyond p53 not detailed","Human relevance of metabolic phenotype not addressed"]},{"year":2026,"claim":"Established a transcriptional input to NUDT16L1, with EGR1 directly activating its promoter to confer oxidative-stress protection in spermatogonial stem cells.","evidence":"ChIP, luciferase reporter assays, overexpression/knockdown with ROS/apoptosis readouts, and a mouse testicular ischemia-reperfusion model","pmids":["41521514"],"confidence":"Medium","gaps":["Single lab","Downstream effectors connecting NUDT16L1 to ROS reduction not defined"]},{"year":null,"claim":"How TIRR's distinct activities—53BP1/p53 inhibition, mRNA export, and mitochondrial NAD-RNA binding—are integrated and switched between under different cellular states remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unifying model linking nuclear and mitochondrial functions","Hierarchy among competing dissociation mechanisms (hairpin RNA, NEAT1_1, ubiquitination) unestablished"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0003723","term_label":"RNA binding","supporting_discovery_ids":[2,5,6,8,9]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,1,3,4]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[0,7]},{"term_id":"GO:0005739","term_label":"mitochondrion","supporting_discovery_ids":[9]}],"pathway":[{"term_id":"R-HSA-73894","term_label":"DNA Repair","supporting_discovery_ids":[0,3,7]},{"term_id":"R-HSA-8953854","term_label":"Metabolism of RNA","supporting_discovery_ids":[6,8]}],"complexes":[],"partners":["TP53BP1","TP53","DTX3L","XPO1","NEAT1","TARDBP","EGR1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9BRJ7","full_name":"Tudor-interacting repair regulator protein","aliases":["NUDT16-like protein 1","Protein syndesmos"],"length_aa":211,"mass_kda":23.3,"function":"Key regulator of TP53BP1 required to stabilize TP53BP1 and regulate its recruitment to chromatin (PubMed:28241136). In absence of DNA damage, interacts with the tandem Tudor-like domain of TP53BP1, masking the region that binds histone H4 dimethylated at 'Lys-20' (H4K20me2), thereby preventing TP53BP1 recruitment to chromatin and maintaining TP53BP1 localization to the nucleus (PubMed:28241136). Following DNA damage, ATM-induced phosphorylation of TP53BP1 and subsequent recruitment of RIF1 leads to dissociate NUDT16L1/TIRR from TP53BP1, unmasking the tandem Tudor-like domain and allowing recruitment of TP53BP1 to DNA double strand breaks (DSBs) (PubMed:28241136). Binds U8 snoRNA (PubMed:18820299)","subcellular_location":"Nucleus","url":"https://www.uniprot.org/uniprotkb/Q9BRJ7/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/NUDT16L1","classification":"Not Classified","n_dependent_lines":214,"n_total_lines":1208,"dependency_fraction":0.1771523178807947},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"CAPZB","stoichiometry":0.2},{"gene":"HNRNPH1","stoichiometry":0.2},{"gene":"UBA1","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/NUDT16L1","total_profiled":1310},"omim":[{"mim_id":"617338","title":"NUDIX HYDROLASE 16-LIKE 1; NUDT16L1","url":"https://www.omim.org/entry/617338"},{"mim_id":"602505","title":"PAXILLIN; PXN","url":"https://www.omim.org/entry/602505"},{"mim_id":"600017","title":"SYNDECAN 4; SDC4","url":"https://www.omim.org/entry/600017"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/NUDT16L1"},"hgnc":{"alias_symbol":["SDOS","TIRR"],"prev_symbol":[]},"alphafold":{"accession":"Q9BRJ7","domains":[{"cath_id":"3.90.79.10","chopping":"9-148","consensus_level":"high","plddt":95.3681,"start":9,"end":148}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BRJ7","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BRJ7-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BRJ7-F1-predicted_aligned_error_v6.png","plddt_mean":93.5},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=NUDT16L1","jax_strain_url":"https://www.jax.org/strain/search?query=NUDT16L1"},"sequence":{"accession":"Q9BRJ7","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9BRJ7.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9BRJ7/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BRJ7"}},"corpus_meta":[{"pmid":"28241136","id":"PMC_28241136","title":"TIRR regulates 53BP1 by masking its histone methyl-lysine binding function.","date":"2017","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/28241136","citation_count":112,"is_preprint":false},{"pmid":"29844495","id":"PMC_29844495","title":"Structural basis for recognition of 53BP1 tandem Tudor domain by TIRR.","date":"2018","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/29844495","citation_count":44,"is_preprint":false},{"pmid":"29967538","id":"PMC_29967538","title":"Mechanism of 53BP1 activity regulation by RNA-binding TIRR and a designer protein.","date":"2018","source":"Nature structural & molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/29967538","citation_count":41,"is_preprint":false},{"pmid":"33961797","id":"PMC_33961797","title":"TIRR inhibits the 53BP1-p53 complex to alter cell-fate programs.","date":"2021","source":"Molecular 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TIRR/53BP1 complex to maintain genome integrity.","date":"2024","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/39349456","citation_count":10,"is_preprint":false},{"pmid":"39632881","id":"PMC_39632881","title":"DTX3L-mediated TIRR nuclear export and degradation regulates DNA repair pathway choice and PARP inhibitor sensitivity.","date":"2024","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/39632881","citation_count":9,"is_preprint":false},{"pmid":"39063270","id":"PMC_39063270","title":"Hypoglycemic Ability of Sericin-Derived Oligopeptides (SDOs) from Bombyx mori Yellow Silk Cocoons and Their Physiological Effects on Streptozotocin (STZ)-Induced Diabetic Rats.","date":"2024","source":"Foods (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/39063270","citation_count":6,"is_preprint":false},{"pmid":"39517289","id":"PMC_39517289","title":"Oral Toxicity and Hypotensive Influence of Sericin-Derived Oligopeptides (SDOs) from Yellow Silk Cocoons of Bombyx mori in Rodent Studies.","date":"2024","source":"Foods (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/39517289","citation_count":5,"is_preprint":false},{"pmid":"32036573","id":"PMC_32036573","title":"TIRR: a potential front runner in HDR race-hypotheses and perspectives.","date":"2020","source":"Molecular biology reports","url":"https://pubmed.ncbi.nlm.nih.gov/32036573","citation_count":4,"is_preprint":false},{"pmid":"39942094","id":"PMC_39942094","title":"Pilot-Scale Production of Sericin-Derived Oligopeptides (SDOs) from Yellow Silk Cocoons: Peptide Characterization and Specifications.","date":"2025","source":"Foods (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/39942094","citation_count":4,"is_preprint":false},{"pmid":"38861384","id":"PMC_38861384","title":"Endogenous p53 inhibitor TIRR dissociates systemic metabolic health from oncogenic activity.","date":"2024","source":"Cell 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research","url":"https://pubmed.ncbi.nlm.nih.gov/39119906","citation_count":1,"is_preprint":false},{"pmid":"41521514","id":"PMC_41521514","title":"Protective Role of the EGR1-Nudt16L1 Pathway in Spermatogonial Stem Cells Against Testicular Ischemia-Reperfusion Injury.","date":"2026","source":"Cell biology international","url":"https://pubmed.ncbi.nlm.nih.gov/41521514","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":10014,"output_tokens":3398,"usd":0.040506,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11043,"output_tokens":3634,"usd":0.073033,"stage2_stop_reason":"end_turn"},"total_usd":0.113539,"stage1_batch_id":"msgbatch_013a8chtv3AWyTqv4z4sEF4C","stage2_batch_id":"msgbatch_01AHsuW63vinsvuTgzmQ3n62","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2017,\n      \"finding\": \"TIRR (NUDT16L1) directly binds the tandem Tudor domain of 53BP1 and masks its H4K20me2 histone methyl-lysine binding motif, preventing 53BP1 recruitment to DNA double-strand breaks. Upon DNA damage, ATM phosphorylates 53BP1 and recruits RIF1 to dissociate the 53BP1-TIRR complex. Overexpression of TIRR impedes 53BP1 localization to DSBs, while depletion of TIRR destabilizes 53BP1 in the nuclear-soluble fraction.\",\n      \"method\": \"Co-immunoprecipitation, overexpression/depletion functional assays, live-cell imaging of DSB foci\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, multiple orthogonal functional assays, replicated by multiple subsequent independent labs\",\n      \"pmids\": [\"28241136\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Crystal structure (1.76 Å) of TIRR in complex with the 53BP1 tandem Tudor domain reveals that the N-terminal region (residues 10–24) and the L8-loop of TIRR interact with 53BP1 Tudor through three loops (L1, L3, L1'), blocking the H4K20me2-binding surface. A TIRR-specific histidine (H106), absent from the homolog NUDT16, is essential for 53BP1 Tudor binding; mutations mimicking TIRR binding modules restore disrupted NUDT16-53BP1 Tudor interaction.\",\n      \"method\": \"X-ray crystallography, site-directed mutagenesis, binding assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — high-resolution crystal structure with mutagenesis validation, independently corroborated by two contemporaneous structural studies\",\n      \"pmids\": [\"29844495\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"X-ray crystal structure of TIRR bound to 53BP1 tandem Tudor domain reveals that an essential TIRR arginine residue is central to an intricate binding area that blocks the methylated-chromatin-binding surface of 53BP1. A 53BP1 separation-of-function mutation abolishing TIRR-mediated regulation renders 53BP1 hyperactive at DSBs. TIRR-interacting RNA molecules relieve this inhibition, providing proof-of-principle of RNA-triggered 53BP1 recruitment to DSBs.\",\n      \"method\": \"X-ray crystallography, site-directed mutagenesis, cell-based DSB foci assays, RNA binding assays\",\n      \"journal\": \"Nature structural & molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure with mutagenesis and functional cell-based validation in one study, corroborated by independent structural studies\",\n      \"pmids\": [\"29967538\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Crystal structure of TIRR–53BP1 tandem Tudor domain complex shows three TIRR loops masking the methylated lysine-binding pocket of 53BP1 TTD, competing with histone H4K20 methylation. Key interaction residues were mapped and their mutation abolishes complex formation. NUDT16 does not directly interact with 53BP1 due to absence of key binding residues. TIRR suppresses relocation of 53BP1 to DNA lesions and 53BP1-dependent DNA damage repair.\",\n      \"method\": \"X-ray crystallography, site-directed mutagenesis, Co-IP, DSB repair assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure with mutagenesis and functional validation; explicitly demonstrates NUDT16 lacks key binding residues as a negative control\",\n      \"pmids\": [\"30002377\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"TIRR inhibits complex formation between the Tudor domain of 53BP1 and dimethylated p53 (K382me2), thereby suppressing p53 transcriptional activation of target genes. Loss of TIRR causes an aberrant increase in p53 gene transactivation, affecting p53-mediated cell-fate programs. TIRR depletion is selectively not tolerated in p53-proficient tumors.\",\n      \"method\": \"Biochemical binding assays (Tudor domain–K382me2 p53 interaction), gene expression analysis upon TIRR loss, cell viability assays\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — biochemical complex reconstitution with cellular functional readout, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"33961797\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RNA with a hairpin secondary structure transcribed at DSBs by RNA polymerase II promotes TIRR/53BP1 complex dissociation. This hairpin RNA binds to the same residues on TIRR as 53BP1, providing the mechanistic basis for RNA-driven complex separation after DNA damage.\",\n      \"method\": \"RNA binding assays, mutagenesis mapping of RNA-TIRR interaction surface, cell-based TIRR/53BP1 dissociation assays, RNA polymerase II inhibition\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct RNA–protein binding mapped to specific residues with functional complex dissociation assay, single lab\",\n      \"pmids\": [\"36288694\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"NEAT1 long non-coding RNA (specifically the short isoform NEAT1_1, enriched in G1 phase) is the primary RNA partner of TIRR within cells, identified by iCLIP. NEAT1_1 binding destabilizes the TIRR/53BP1 complex, promoting 53BP1 function in a cell-cycle-dependent manner. TDP-43 modulates the TIRR/53BP1 complex by promoting NEAT1_1 production.\",\n      \"method\": \"iCLIP (individual-nucleotide resolution UV crosslinking and immunoprecipitation), Co-IP, cell-cycle synchronization, TDP-43 functional assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — iCLIP identifies direct RNA–protein contacts; functional disruption of TIRR/53BP1 complex validated, single lab\",\n      \"pmids\": [\"39349456\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"DTX3L ubiquitinates TIRR at lysine 187, facilitating XPO1-mediated nuclear export and subsequent degradation of TIRR upon DNA damage. This relieves TIRR-mediated inhibition of 53BP1, regulating NHEJ pathway activity and PARP inhibitor sensitivity. DTX3L overexpression (as in prostate cancers) decreases TIRR levels, impairs HR, and induces chromosomal instability.\",\n      \"method\": \"Ubiquitination assays, nuclear export inhibition (XPO1 inhibitor), site-directed mutagenesis (K187), co-immunoprecipitation, cellular fractionation, PARP inhibitor sensitivity assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — specific ubiquitination site identified with mutagenesis, export mechanism validated with inhibitor, multiple orthogonal functional readouts\",\n      \"pmids\": [\"39632881\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"TIRR selectively binds a subset of mRNAs in response to DNA damage and interacts with the nuclear export protein Exportin-1 (XPO1/CRM1) through a nuclear export signal. TIRR and TIRR-bound RNA co-localize with processing bodies (P-bodies); TIRR depletion results in nuclear RNA retention and impaired P-body formation, linking TIRR's RNA-binding activity to mRNA nuclear export and storage during the DDR.\",\n      \"method\": \"RNA immunoprecipitation, co-immunoprecipitation with XPO1, fluorescence microscopy (P-body co-localization), TIRR depletion with mRNA export/P-body formation readouts\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — direct protein–RNA and protein–XPO1 interactions demonstrated, P-body role validated by depletion, single lab single study\",\n      \"pmids\": [\"39119906\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"NUDT16L1 localizes to mitochondria in colon cancer cells, where it prevents mitochondrial DNA leakage upon ferroptosis induction. It promotes ferroptosis insensitivity by binding directly to NAD-capped RNAs and indirectly enhancing expression of ferroptosis repressor and mitochondrial genes through MALAT1.\",\n      \"method\": \"Subcellular fractionation/mitochondrial localization, NAD-capped RNA binding assays, NUDT16L1 overexpression/knockdown with ferroptosis assays, transgenic mouse model\",\n      \"journal\": \"Redox biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct mitochondrial localization with functional consequence (mtDNA leakage), direct NAD-capped RNA binding demonstrated, in vivo model corroboration\",\n      \"pmids\": [\"39317106\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Deletion of TIRR in mice selectively activates p53, protecting against cancer but causing systemic metabolic imbalance (overweight, insulin resistance). These metabolic and oncoprotective effects are dependent on p53. Tissue-specific models indicate glucose homeostasis is regulated primarily by TIRR expression in adipose tissue, and orexigenesis by TIRR expression in the CNS.\",\n      \"method\": \"Conditional/tissue-specific mouse knockout models, metabolic phenotyping, p53 deletion epistasis\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — tissue-specific KO with epistasis to p53 establishes pathway placement; in vivo mouse models with defined metabolic phenotypes\",\n      \"pmids\": [\"38861384\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"EGR1 transcription factor directly binds the promoter of NUDT16L1 and activates its expression, establishing NUDT16L1 as a downstream effector of EGR1. Overexpression of NUDT16L1 alone reduces ROS and prevents cell death in spermatogonial stem cells under oxidative stress; silencing NUDT16L1 abolishes EGR1's protective effects against testicular ischemia-reperfusion injury in mice.\",\n      \"method\": \"Chromatin immunoprecipitation (ChIP), luciferase reporter assay, NUDT16L1 overexpression/knockdown with ROS/apoptosis readouts, mouse testicular IR injury model\",\n      \"journal\": \"Cell biology international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP and luciferase confirm direct transcriptional regulation; epistasis (KD abolishes EGR1 effect; OE mimics it) places NUDT16L1 downstream, single lab\",\n      \"pmids\": [\"41521514\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"NUDT16L1/TIRR is an RNA-binding protein that directly binds the tandem Tudor domain of 53BP1 (via its N-terminal region, L8-loop, and a critical H106 residue), masking its H4K20me2 and K382me2-p53 binding surfaces to inhibit 53BP1 recruitment to DNA double-strand breaks and 53BP1-mediated p53 activation; upon DNA damage, DSB-derived hairpin RNA and NEAT1_1 lncRNA compete for the same TIRR surface to dissociate the TIRR/53BP1 complex, while DTX3L-mediated ubiquitination of TIRR at K187 drives XPO1-dependent nuclear export and proteolytic degradation of TIRR, collectively relieving 53BP1 inhibition; TIRR additionally functions as an mRNA export regulator via XPO1 and modulates P-body formation, and in certain cancer contexts localizes to mitochondria to bind NAD-capped RNAs and suppress ferroptosis, with the EGR1 transcription factor acting as an upstream activator of NUDT16L1 transcription in spermatogonial stem cells.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"NUDT16L1 (TIRR) is an RNA-binding regulator of the DNA damage response that controls 53BP1 activity by directly binding its tandem Tudor domain [#0]. High-resolution crystal structures show that the TIRR N-terminal region and L8-loop engage 53BP1 Tudor through three loops, masking the methyl-lysine binding surface that 53BP1 otherwise uses to read H4K20me2 chromatin marks; a TIRR-specific histidine (H106) and an essential arginine are central to this interface, and these residues distinguish TIRR from its homolog NUDT16, which does not bind 53BP1 [#1, #2, #3]. By occluding this surface, TIRR prevents 53BP1 recruitment to double-strand breaks and dampens DNA repair, while a parallel interaction blocks 53BP1 binding to dimethylated p53 (K382me2), suppressing p53-dependent transcription; loss of TIRR hyperactivates p53 and is selectively lethal in p53-proficient tumors [#0, #4]. TIRR inhibition is relieved upon DNA damage through competing RNAs that bind the same TIRR surface as 53BP1—DSB-derived hairpin RNAs and the NEAT1_1 lncRNA isoform dissociate the TIRR/53BP1 complex in a cell-cycle-dependent manner [#2, #5, #6]—and through DTX3L-mediated ubiquitination of TIRR at K187, which drives XPO1-dependent nuclear export and degradation [#7]. Beyond 53BP1 regulation, TIRR couples to mRNA metabolism, binding selected mRNAs and the export factor XPO1 to direct nuclear RNA export and P-body formation during the DDR [#8]. In mice, TIRR loss activates p53 to confer cancer protection at the cost of systemic metabolic imbalance [#10]. In specific cancer contexts NUDT16L1 localizes to mitochondria, binds NAD-capped RNAs, and suppresses ferroptosis [#9], and its transcription is directly activated by EGR1 to protect spermatogonial stem cells from oxidative stress [#11].\",\n  \"teleology\": [\n    {\n      \"year\": 2017,\n      \"claim\": \"Established that TIRR is a direct negative regulator of 53BP1, answering how 53BP1 recruitment to breaks is held in check before damage.\",\n      \"evidence\": \"Co-IP, overexpression/depletion functional assays, and live-cell DSB foci imaging in human cells\",\n      \"pmids\": [\"28241136\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not define the atomic basis of the TIRR/53BP1 interface\", \"Mechanism of RNA-driven dissociation not yet established\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Crystal structures resolved how TIRR masks the 53BP1 Tudor methyl-lysine pocket and identified the specific residues (H106, an essential arginine) that confer specificity, explaining why the homolog NUDT16 cannot bind.\",\n      \"evidence\": \"X-ray crystallography of TIRR-53BP1 Tudor complexes with site-directed mutagenesis, DSB foci and repair assays, and RNA binding assays across three independent studies\",\n      \"pmids\": [\"29844495\", \"29967538\", \"30002377\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the physiological dissociating RNA species in cells not yet defined\", \"How damage signaling triggers dissociation in vivo not fully resolved\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Extended TIRR's regulatory reach beyond chromatin to p53, showing it blocks the 53BP1 Tudor-K382me2-p53 interaction and thereby restrains p53 transactivation, defining a synthetic vulnerability in p53-proficient tumors.\",\n      \"evidence\": \"Biochemical Tudor-K382me2 binding assays, gene expression analysis on TIRR loss, and cell viability assays\",\n      \"pmids\": [\"33961797\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Cell-type specificity of the p53 axis not fully mapped\", \"Did not address how this is coordinated with the DSB-recruitment function\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Identified the molecular trigger for relieving TIRR inhibition after damage: DSB-transcribed hairpin RNAs bind the same TIRR residues as 53BP1 and competitively dissociate the complex.\",\n      \"evidence\": \"RNA binding assays, mutagenesis mapping of the RNA-TIRR surface, RNA Pol II inhibition, and cell-based dissociation assays\",\n      \"pmids\": [\"36288694\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab; reciprocal validation of the in-cell hairpin species limited\", \"Kinetics relative to RIF1/ATM-driven dissociation not resolved\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Defined NEAT1_1 lncRNA as the primary cellular RNA partner of TIRR and linked its TDP-43-dependent production to cell-cycle-dependent control of 53BP1.\",\n      \"evidence\": \"iCLIP, Co-IP, cell-cycle synchronization, and TDP-43 functional assays\",\n      \"pmids\": [\"39349456\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab; relationship between NEAT1_1 and DSB hairpin RNA competition not reconciled\", \"Quantitative contribution of TDP-43 in vivo not established\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Revealed a degradative arm of TIRR regulation, showing DTX3L ubiquitinates TIRR at K187 to drive XPO1-dependent nuclear export and degradation, coupling TIRR turnover to repair-pathway choice and PARP inhibitor sensitivity.\",\n      \"evidence\": \"Ubiquitination assays, K187 mutagenesis, XPO1 inhibition, fractionation, and PARP inhibitor sensitivity assays\",\n      \"pmids\": [\"39632881\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How RNA-driven and ubiquitination-driven dissociation are temporally coordinated unclear\", \"Generality of DTX3L overexpression effect across cancer types not fully tested\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Expanded TIRR function to mRNA metabolism, demonstrating XPO1-coupled mRNA export and P-body involvement during the DDR.\",\n      \"evidence\": \"RNA-IP, Co-IP with XPO1, fluorescence microscopy of P-body co-localization, and depletion readouts of RNA export\",\n      \"pmids\": [\"39119906\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab single study\", \"Identity and fate of the exported mRNA cargo not defined\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identified a context-specific mitochondrial role for NUDT16L1 in binding NAD-capped RNAs and suppressing ferroptosis in colon cancer.\",\n      \"evidence\": \"Subcellular fractionation, NAD-capped RNA binding assays, ferroptosis assays, and a transgenic mouse model\",\n      \"pmids\": [\"39317106\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanistic link to the nuclear 53BP1/RNA functions unclear\", \"Generality beyond colon cancer not established\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Placed TIRR upstream of organismal phenotypes via p53, showing tissue-specific knockouts produce cancer protection alongside metabolic imbalance in a p53-dependent manner.\",\n      \"evidence\": \"Conditional/tissue-specific mouse knockouts, metabolic phenotyping, and p53 deletion epistasis\",\n      \"pmids\": [\"38861384\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular link between TIRR and tissue metabolism beyond p53 not detailed\", \"Human relevance of metabolic phenotype not addressed\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Established a transcriptional input to NUDT16L1, with EGR1 directly activating its promoter to confer oxidative-stress protection in spermatogonial stem cells.\",\n      \"evidence\": \"ChIP, luciferase reporter assays, overexpression/knockdown with ROS/apoptosis readouts, and a mouse testicular ischemia-reperfusion model\",\n      \"pmids\": [\"41521514\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab\", \"Downstream effectors connecting NUDT16L1 to ROS reduction not defined\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How TIRR's distinct activities—53BP1/p53 inhibition, mRNA export, and mitochondrial NAD-RNA binding—are integrated and switched between under different cellular states remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unifying model linking nuclear and mitochondrial functions\", \"Hierarchy among competing dissociation mechanisms (hairpin RNA, NEAT1_1, ubiquitination) unestablished\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003723\", \"supporting_discovery_ids\": [2, 5, 6, 8, 9]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 1, 3, 4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0, 7]},\n      {\"term_id\": \"GO:0005739\", \"supporting_discovery_ids\": [9]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-73894\", \"supporting_discovery_ids\": [0, 3, 7]},\n      {\"term_id\": \"R-HSA-8953854\", \"supporting_discovery_ids\": [6, 8]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"TP53BP1\", \"TP53\", \"DTX3L\", \"XPO1\", \"NEAT1\", \"TARDBP\", \"EGR1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}