{"gene":"CRISP3","run_date":"2026-04-28T17:28:53","timeline":{"discoveries":[{"year":1996,"finding":"CRISP3 (SGP28) is localized in specific granules of human neutrophils, as determined by subcellular fractionation of human neutrophils and partial primary structure determination of the purified 28 kDa glycoprotein.","method":"Protein purification from neutrophil exocytosed material, cDNA cloning, subcellular fractionation","journal":"FEBS letters","confidence":"High","confidence_rationale":"Tier 2 — direct subcellular fractionation with protein purification and sequencing, replicated in subsequent studies","pmids":["8601434"],"is_preprint":false},{"year":2002,"finding":"CRISP3 is a matrix protein localized in a subset of peroxidase-negative granules with overlapping characteristics of specific and gelatinase granules in neutrophils, and is also present in granules of eosinophils; it exists as both glycosylated and unglycosylated forms with identical subcellular distribution.","method":"Three-layer Percoll density gradient fractionation, secretagogue release studies, double-labeling immunogold electron microscopy","journal":"Journal of leukocyte biology","confidence":"High","confidence_rationale":"Tier 1–2 — multiple orthogonal methods including immunogold EM and subcellular fractionation with functional mobilization studies","pmids":["12223513"],"is_preprint":false},{"year":2005,"finding":"Beta-microseminoprotein (MSP/PSP94) and CRISP3 form stable, non-covalent high-affinity complexes in human seminal plasma, with CRISP3 binding MSP through its amino-terminal SCP-domain.","method":"Immunoprecipitation, gel filtration, surface plasmon resonance","journal":"Biochemical and biophysical research communications","confidence":"High","confidence_rationale":"Tier 1–2 — reciprocal Co-IP, gel filtration, and quantitative SPR binding kinetics in a single study","pmids":["15950934"],"is_preprint":false},{"year":1996,"finding":"CRISP3 expression in mouse B cells is transcriptionally activated by the Oct2 transcription factor via two variant octamer motifs in the CRISP3 promoter; the C-terminal transactivation domain of Oct2 is required, and CRISP3 is specifically expressed at the pre-B-cell stage.","method":"Nuclear run-on experiments, Oct2-deficient primary B cells, cotransfection reporter assays, site-directed mutagenesis of octamer motifs, in vitro Oct2–DNA binding","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 1–2 — multiple orthogonal methods including nuclear run-on, knockout primary cells, and mutagenesis of regulatory elements","pmids":["8887646"],"is_preprint":false},{"year":1993,"finding":"CRISP3 gene expression in mouse salivary gland is strongly androgen-dependent, as demonstrated by castration abolishing/reducing expression in male mice and absence of expression in female salivary gland.","method":"cDNA cloning, RNA blot analysis of castrated vs. intact males and females","journal":"Endocrinology","confidence":"Medium","confidence_rationale":"Tier 2 — direct hormonal manipulation with quantitative RNA blotting, single lab","pmids":["8319566"],"is_preprint":false},{"year":1995,"finding":"The mouse CRISP3 gene spans over 20 kb, consists of eight exons and seven introns, has a TATA box-proximal transcription start site, and contains two androgen-responsive element consensus sequences in its promoter along with putative OTF- and GATA-binding elements.","method":"Genomic library screening, DNA sequencing, primer extension mapping, PCR sizing of introns","journal":"The Biochemical journal","confidence":"Medium","confidence_rationale":"Tier 2 — direct genomic characterization with promoter element identification","pmids":["7639699"],"is_preprint":false},{"year":2008,"finding":"NMR analysis of the MSP–CRISP3 complex shows that only one side of MSP (comprising beta-strands 1, 4, 5, and 8, with beta-strands 1 and 8 forming the main binding surface) is affected by complex formation with the N-terminal SCP domain of CRISP3.","method":"Multidimensional NMR (15N-HSQC and 3D-NMR of triply-labeled MSP in complex with recombinant N-terminal CRISP3 domain)","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 1 — NMR structural characterization of the complex, but model is tentative (no full CRISP3 structure)","pmids":["19026612"],"is_preprint":false},{"year":2011,"finding":"Equine CRISP3 purified from seminal plasma suppresses binding between spermatozoa and polymorphonuclear neutrophils (PMNs), identifying CRISP3 as a seminal plasma protein that modulates sperm elimination from the female reproductive tract.","method":"Sequential ammonium sulfate precipitation, size-exclusion and ion-exchange chromatography, 1D/2D SDS-PAGE, Western blotting, flow cytometry PMN/sperm binding assay","journal":"Biology of reproduction","confidence":"High","confidence_rationale":"Tier 2 — protein purified to homogeneity, functional activity confirmed by flow cytometry with appropriate controls","pmids":["21389342"],"is_preprint":false},{"year":2010,"finding":"Human CRISP3 binds alpha-1B-glycoprotein (A1BG) in serum; A1BG orthologs from cow, horse, and rabbit also bind CRISP3, while mouse kininogen-1 binds CRISP3 in mouse serum, suggesting a conserved mechanism of CRISP binding by A1BG-family proteins.","method":"Gel filtration, ligand blotting, affinity isolation, mass spectrometry, N-terminal sequencing","journal":"Biochimica et biophysica acta","confidence":"Medium","confidence_rationale":"Tier 2 — affinity isolation with MS identification across multiple species, single lab","pmids":["20116414"],"is_preprint":false},{"year":2013,"finding":"The binding interface of PSP94–CRISP3 complex was mapped: PSP94 residues Y3, F4, P56, and its C-terminal beta-strand (plus the C37–C73 disulfide bond) are critical for CRISP3 binding; on CRISP3, the N-terminal SCP domain alone is insufficient—the hinge region is also required, but the C-terminal ICR domain is not.","method":"Site-directed mutagenesis of PSP94 and CRISP3, co-immunoprecipitation","journal":"Biochimica et biophysica acta","confidence":"Medium","confidence_rationale":"Tier 2 — mutagenesis combined with Co-IP to map binding residues, single lab","pmids":["23375721"],"is_preprint":false},{"year":2014,"finding":"Native, glycosylated human and mouse CRISP3 expressed in HEK 293 cells exist as monomers in solution; N-glycosylation sites and patterns differ between human and mouse CRISP3.","method":"Eukaryotic expression in HEK 293 cells, ion exchange and size exclusion chromatography, substrate-affinity assays, glycosylation characterization","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 — native protein purification with functional validation, single lab","pmids":["24573035"],"is_preprint":false},{"year":2011,"finding":"CRISP3 is a direct transcriptional target of the ERG transcription factor (driven by TMPRSS2-ERG fusion) in prostate cancer, as shown by chromatin immunoprecipitation with an anti-ERG antibody demonstrating ERG binding at the CRISP3 locus.","method":"Genome-wide mRNA expression analysis, quantitative RT-PCR, immunohistochemistry, chromatin immunoprecipitation (ChIP) with anti-ERG antibody","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 — ChIP directly demonstrates ERG occupancy at CRISP3 locus, supported by expression correlation","pmids":["21814574"],"is_preprint":false},{"year":2018,"finding":"CRISP3 expression in prostate cancer cells is androgen-dependent and regulated by androgen receptor (AR) through epigenetic mechanisms: in AR-negative cells the CRISP3 promoter is silenced by histone deacetylation; DHT treatment of LNCaP cells increases CRISP3 transcript and protein; AR occupancy at the CRISP3 promoter was confirmed by ChIP.","method":"Luciferase reporter assays with CRISP3 promoter fragments, ChIP-PCR, DHT treatment, histone deacetylase inhibition","journal":"The Journal of steroid biochemistry and molecular biology","confidence":"Medium","confidence_rationale":"Tier 2 — multiple orthogonal methods (reporter assay, ChIP, hormone treatment) in a single lab","pmids":["29477539"],"is_preprint":false},{"year":2015,"finding":"Recombinant CRISP3 enhances adhesion and proliferation of human endometrial epithelial cells in vitro; CRISP3 is secreted by primary human endometrial epithelial cells and accumulates in uterine lavage fluid, with higher abundance during the proliferative phase.","method":"Treatment of endometrial epithelial cells with recombinant CRISP3, adhesion and proliferation assays, uterine lavage ELISA, immunohistochemistry","journal":"Biology of reproduction","confidence":"Medium","confidence_rationale":"Tier 2 — recombinant protein functional assay with defined cellular readouts, single lab","pmids":["25715794"],"is_preprint":false},{"year":2020,"finding":"CRISP3 facilitates prostate cancer progression from carcinoma in situ to invasive cancer in vivo; CRISP3 exposure enhances cancer cell motility and invasion; mass spectrometry revealed CRISP3 induces changes in abundance of cell-cell adhesion proteins LASP1 and TJP1 both in vivo and in vitro.","method":"Genetically engineered mouse model of prostate cancer, human and mouse prostate cancer cell motility/invasion assays, mass spectrometry proteomics","journal":"Endocrine-related cancer","confidence":"High","confidence_rationale":"Tier 2 — in vivo mouse model combined with cell-based functional assays and proteomics, multiple orthogonal approaches","pmids":["32357309"],"is_preprint":false},{"year":2010,"finding":"PSP94 and CRISP3 each independently inhibit prostate cancer cell growth in a cell-line-specific manner; growth inhibition by CRISP3 is not affected by presence or absence of PSP94, suggesting CRISP3 participates in PSP94-independent activities.","method":"Ectopic expression of PSP94 and CRISP3 in prostate cell lines, clonogenic survival assay, Western blot, immunofluorescence","journal":"Asian journal of andrology","confidence":"Medium","confidence_rationale":"Tier 2 — gain-of-function with defined cellular phenotype, single lab","pmids":["20676114"],"is_preprint":false},{"year":2016,"finding":"CRISP3 was purified from human seminal plasma using PSP94-immobilized affinity chromatography and confirmed to bind PSP94 with high affinity by surface plasmon resonance; it exists as glycosylated (~28 kDa) and unglycosylated (~26 kDa) forms.","method":"PSP94-affinity chromatography, SDS-PAGE, immunoblotting, MALDI-TOF MS, deglycosylation, surface plasmon resonance","journal":"Journal of chromatography B","confidence":"Medium","confidence_rationale":"Tier 2 — protein purified to homogeneity with binding kinetics validated by SPR","pmids":["27825912"],"is_preprint":false},{"year":2024,"finding":"Exogenous CRISP3 downregulates P2RX7 (an ATP-gated ion channel) in PC3 prostate cancer cells and THP1 macrophages, reducing ATP-induced cytotoxicity and IL-1β secretion; this effect is abrogated when CRISP3 is complexed with PSP94. CRISP3 mediates these effects through CITED2 (a transcriptional coregulator that reduces p300 availability at the P2RX7 promoter); PSP94 also affects CRISP3 endocytosis and its interaction with flotillin-2.","method":"Recombinant protein treatment, P2RX7 expression assays, ATP cytotoxicity assay, IL-1β ELISA, antibody array, CITED2 overexpression, CUT&RUN assay for p300 at P2RX7 promoter, Co-IP with flotillin-2","journal":"Biochimica et biophysica acta. Molecular cell research","confidence":"Medium","confidence_rationale":"Tier 2 — multiple orthogonal mechanistic methods in a single lab defining a novel signaling axis","pmids":["40473221"],"is_preprint":false},{"year":2024,"finding":"Boar CRISP3 is localized in the post-acrosomal region of the sperm head and migrates to the anterior end of the tail after capacitation; recombinant CRISP3 downregulates inflammatory factors IL-1α, IL-1β, and IL-6 in LPS-stimulated RAW264.7 macrophages, indicating immunomodulatory function.","method":"Immunofluorescence localization, recombinant protein treatment of macrophages, RT-PCR and Western blot for inflammatory cytokines","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2 — direct localization experiment linked to functional immunomodulatory readout, single lab","pmids":["38396941"],"is_preprint":false},{"year":2024,"finding":"Equine seminal plasma CRISP3 selectively suppresses PMN binding to live spermatozoa but not to dead spermatozoa or bacteria; immunocytochemistry confirmed CRISP3 binds to live but not dead spermatozoa, suggesting selective recognition of viable sperm surface.","method":"CRISP3 purification from seminal plasma, flow cytometry PMN binding assay with live/dead sperm populations, immunocytochemistry","journal":"Theriogenology","confidence":"Medium","confidence_rationale":"Tier 2 — purified protein functional assay with flow cytometry and localization, single lab","pmids":["38377715"],"is_preprint":false},{"year":2024,"finding":"EP300 histone acetyltransferase regulates CRISP3 expression in triple-negative breast cancer by modifying H3K27ac at the CRISP3 promoter; CRISP3 overexpression promotes tumor stemness and lobaplatin resistance, and knockdown of EP300 reduces these phenotypes in a manner rescued by CRISP3 overexpression.","method":"ChIP for H3K27ac at CRISP3 promoter, EP300 knockdown and CRISP3 overexpression rescue experiments, in vitro and in vivo stemness/resistance assays","journal":"Human cell","confidence":"Medium","confidence_rationale":"Tier 2 — ChIP-based epigenetic mechanism with epistatic rescue experiment, single lab","pmids":["38879857"],"is_preprint":false},{"year":2022,"finding":"hsa_circ_0003823 promotes ESCC progression and apatinib resistance through the miR-607/CRISP3 axis: the circular RNA sequesters miR-607, leading to upregulation of CRISP3; CRISP3 was validated as a miR-607 target by RNA immunoprecipitation and dual-luciferase reporter assays.","method":"RNA immunoprecipitation (RIP), dual-luciferase reporter assay, Western blot, knockdown experiments","journal":"International journal of biological sciences","confidence":"Medium","confidence_rationale":"Tier 2 — RIP and luciferase assays directly demonstrate miR-607 targeting of CRISP3, single lab","pmids":["36263172"],"is_preprint":false},{"year":2026,"finding":"CRISP3 acts as a tumor suppressor in high-grade serous ovarian carcinoma by inhibiting cell proliferation, migration, and invasion through suppression of the PI3K/AKT signaling pathway; CRISP3 knockdown activates PI3K/AKT and overexpression suppresses it.","method":"CRISP3 knockdown and overexpression in HGSOC cells, CCK-8/EdU proliferation and Transwell invasion assays, RNA-seq, GSEA, Western blotting for PI3K/AKT pathway components, in vivo mouse model","journal":"Biomedicines","confidence":"Medium","confidence_rationale":"Tier 2 — loss- and gain-of-function with pathway readout by Western blot and RNA-seq, single lab","pmids":["41751369"],"is_preprint":false}],"current_model":"CRISP3 is a secreted, cysteine-rich glycoprotein (with glycosylated and unglycosylated forms) that is stored in neutrophil specific/gelatinase granules and exocrine secretions; its expression is controlled by androgens (via AR-mediated epigenetic regulation), Oct2, ERG (in TMPRSS2-ERG fusion prostate cancer), and EP300-mediated H3K27 acetylation; it forms high-affinity non-covalent complexes with MSP/PSP94 through its N-terminal SCP domain and hinge region, binds serum A1BG, and functionally modulates immune responses by suppressing PMN–sperm binding, downregulating inflammatory cytokines, and reducing P2RX7-mediated IL-1β secretion through a CITED2/p300 axis; it also influences cell adhesion (via LASP1, TJP1), promotes prostate cancer invasion, and suppresses ovarian cancer progression via the PI3K/AKT pathway."},"narrative":{"teleology":[{"year":1993,"claim":"Establishing that CRISP3 is an androgen-regulated gene answered the question of what controls its tissue-specific expression in exocrine glands.","evidence":"Castration and RNA blot analysis in mouse salivary gland","pmids":["8319566"],"confidence":"Medium","gaps":["Single tissue (salivary gland) examined","Androgen-responsive elements in promoter not functionally validated"]},{"year":1996,"claim":"Identification of CRISP3 as a specific granule protein of neutrophils established it as a component of the innate immune arsenal, while independent work showed Oct2-driven transcription in pre-B cells revealed a second lineage of expression.","evidence":"Subcellular fractionation and protein purification from neutrophils; nuclear run-on, Oct2-knockout B cells, and reporter assays in mouse","pmids":["8601434","8887646"],"confidence":"High","gaps":["Function of CRISP3 after neutrophil degranulation unknown","Whether Oct2-driven B-cell expression is conserved in humans"]},{"year":2002,"claim":"Refined granule subtyping showed CRISP3 resides in a subset of peroxidase-negative granules overlapping specific and gelatinase granules, and extended expression to eosinophils, broadening its potential role in granulocyte biology.","evidence":"Three-layer Percoll fractionation, secretagogue mobilization, double-labeling immunogold EM in human neutrophils and eosinophils","pmids":["12223513"],"confidence":"High","gaps":["Functional consequence of CRISP3 release from granulocyte granules not tested","Receptor or target on effector cells unknown"]},{"year":2005,"claim":"Discovery that CRISP3 forms a stable high-affinity complex with PSP94/MSP in seminal plasma via its SCP domain identified a major extracellular binding partner and suggested functional modulation of both proteins.","evidence":"Reciprocal co-immunoprecipitation, gel filtration, and surface plasmon resonance in human seminal plasma","pmids":["15950934"],"confidence":"High","gaps":["Biological consequence of complex formation unknown","Binding stoichiometry in vivo not established"]},{"year":2008,"claim":"NMR mapping of the PSP94–CRISP3 interface revealed which PSP94 surfaces are engaged, providing the first structural picture of CRISP3 complex formation.","evidence":"Multidimensional NMR of 15N/13C/2H-labeled PSP94 in complex with recombinant CRISP3 N-terminal domain","pmids":["19026612"],"confidence":"Medium","gaps":["Full CRISP3 structure not determined","NMR model tentative without full-length complex structure","Hinge region contribution not captured"]},{"year":2010,"claim":"Two advances clarified CRISP3's broader interaction network and its cancer-cell-autonomous activity: identification of A1BG as a serum carrier across species, and demonstration that CRISP3 can independently inhibit prostate cancer cell growth.","evidence":"Gel filtration/affinity isolation/MS for A1BG binding; ectopic expression and clonogenic assays in prostate cancer cells","pmids":["20116414","20676114"],"confidence":"Medium","gaps":["Functional consequence of A1BG–CRISP3 interaction unknown","Mechanism of growth inhibition undefined","Both findings from single laboratories"]},{"year":2011,"claim":"Two studies established CRISP3's roles in reproduction and oncogenic transcription: equine CRISP3 suppresses PMN–sperm binding, protecting spermatozoa, while ERG directly transactivates CRISP3 in TMPRSS2-ERG fusion prostate cancer.","evidence":"Purified protein flow cytometry PMN/sperm binding assay; ChIP with anti-ERG antibody at CRISP3 locus in prostate cancer","pmids":["21389342","21814574"],"confidence":"High","gaps":["CRISP3 receptor on PMNs or sperm not identified","Whether ERG-driven CRISP3 is functionally required for prostate cancer phenotype not tested"]},{"year":2013,"claim":"Detailed mutagenesis of both PSP94 and CRISP3 established that the SCP domain plus hinge region of CRISP3 (but not ICR) and specific PSP94 residues are critical for complex formation, refining the structural model.","evidence":"Site-directed mutagenesis combined with co-immunoprecipitation","pmids":["23375721"],"confidence":"Medium","gaps":["No crystal structure of the complex","Functional significance of each binding-site residue untested in vivo"]},{"year":2018,"claim":"AR-mediated epigenetic regulation of CRISP3 in prostate cancer was demonstrated, showing AR occupancy at the CRISP3 promoter and that histone deacetylation silences CRISP3 in AR-negative cells.","evidence":"ChIP-PCR for AR, luciferase reporters, DHT treatment and HDAC inhibition in LNCaP cells","pmids":["29477539"],"confidence":"Medium","gaps":["Specific histone marks at the CRISP3 promoter not mapped in this study","Relationship between AR and ERG-driven regulation not addressed"]},{"year":2020,"claim":"An in vivo prostate cancer model demonstrated that CRISP3 promotes progression from carcinoma in situ to invasive cancer, mechanistically linked to altered abundance of cell-adhesion proteins LASP1 and TJP1.","evidence":"Genetically engineered mouse model, cell motility/invasion assays, mass spectrometry proteomics","pmids":["32357309"],"confidence":"High","gaps":["Direct interaction of CRISP3 with LASP1 or TJP1 not shown","Whether CRISP3 acts intracellularly or extracellularly to modulate adhesion proteins unclear"]},{"year":2024,"claim":"Multiple 2024 studies collectively expanded the functional repertoire: CRISP3 downregulates P2RX7 via CITED2/p300 to suppress IL-1β secretion (blocked when complexed with PSP94); boar CRISP3 downregulates inflammatory cytokines in macrophages; equine CRISP3 selectively protects live sperm from PMN attack; and EP300-mediated H3K27ac drives CRISP3 expression that promotes stemness and drug resistance in triple-negative breast cancer.","evidence":"CUT&RUN for p300 at P2RX7, Co-IP with flotillin-2, macrophage cytokine assays, flow cytometry with live/dead sperm, ChIP for H3K27ac at CRISP3 promoter with epistatic rescue in TNBC","pmids":["40473221","38396941","38377715","38879857"],"confidence":"Medium","gaps":["CITED2/p300 axis validated in one cell system only","Surface receptor mediating CRISP3 signaling into target cells unidentified","EP300/CRISP3 axis in breast cancer not independently replicated"]},{"year":2026,"claim":"CRISP3 was shown to function as a tumor suppressor in high-grade serous ovarian carcinoma by inhibiting PI3K/AKT signaling, contrasting with its pro-invasive role in prostate cancer and revealing context-dependent oncogenic behavior.","evidence":"Loss- and gain-of-function in HGSOC cells, RNA-seq/GSEA, PI3K/AKT pathway Western blotting, in vivo mouse xenograft","pmids":["41751369"],"confidence":"Medium","gaps":["Single-lab finding not independently replicated","Mechanism by which CRISP3 suppresses PI3K/AKT not defined","Contradiction with pro-tumorigenic roles in other cancers unexplained"]},{"year":null,"claim":"The cell-surface receptor(s) through which extracellular CRISP3 signals remain unidentified, and there is no high-resolution structure of full-length CRISP3 or its complexes.","evidence":"","pmids":[],"confidence":"High","gaps":["No receptor identified on neutrophils, sperm, or cancer cells","No crystal or cryo-EM structure of full-length CRISP3","Context-dependent tumor-promoting versus tumor-suppressing activities mechanistically unexplained"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[7,17,19,22]},{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[13,14,17,18]}],"localization":[{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[0,1]},{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[2,7,8,16,19]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[0,1,7,17,18,19]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[17,22]},{"term_id":"R-HSA-1474165","term_label":"Reproduction","supporting_discovery_ids":[7,19]}],"complexes":["PSP94-CRISP3 complex","A1BG-CRISP3 complex"],"partners":["MSMB","A1BG","CITED2","EP300","FLOT2","LASP1","TJP1","P2RX7"],"other_free_text":[]},"mechanistic_narrative":"CRISP3 is a secreted cysteine-rich glycoprotein that functions at the intersection of innate immunity, reproductive biology, and cancer progression. It is stored in specific and gelatinase granules of neutrophils and eosinophils, from which it is released upon activation, and is also secreted by exocrine glands and endometrial epithelium [PMID:8601434, PMID:12223513, PMID:25715794]. CRISP3 forms high-affinity non-covalent complexes with PSP94/MSP through its N-terminal SCP domain and hinge region, and binds the serum carrier alpha-1B-glycoprotein (A1BG); the PSP94 interaction modulates CRISP3 activity, including its ability to downregulate P2RX7 expression via a CITED2/p300 transcriptional axis, thereby suppressing IL-1β secretion [PMID:15950934, PMID:23375721, PMID:20116414, PMID:40473221]. Its expression is controlled by androgens via AR-mediated promoter occupancy, by the ERG transcription factor in TMPRSS2-ERG fusion-positive prostate cancers, and by EP300-dependent H3K27 acetylation; functionally, CRISP3 suppresses PMN–sperm binding to protect viable spermatozoa, downregulates inflammatory cytokines in macrophages, promotes prostate cancer invasion by altering cell-adhesion proteins LASP1 and TJP1, and suppresses ovarian cancer progression through inhibition of PI3K/AKT signaling [PMID:29477539, PMID:21814574, PMID:38879857, PMID:21389342, PMID:38396941, PMID:32357309, PMID:41751369]."},"prefetch_data":{"uniprot":{"accession":"P54108","full_name":"Cysteine-rich secretory protein 3","aliases":["Specific granule protein of 28 kDa","SGP28"],"length_aa":245,"mass_kda":27.6,"function":"","subcellular_location":"Secreted","url":"https://www.uniprot.org/uniprotkb/P54108/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/CRISP3","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/CRISP3","total_profiled":1310},"omim":[{"mim_id":"618062","title":"CYSTEINE-RICH SECRETORY PROTEIN 3; CRISP3","url":"https://www.omim.org/entry/618062"},{"mim_id":"157145","title":"MICROSEMINOPROTEIN, BETA; MSMB","url":"https://www.omim.org/entry/157145"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Group enriched","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"fallopian tube","ntpm":1133.0},{"tissue":"salivary gland","ntpm":2263.8}],"url":"https://www.proteinatlas.org/search/CRISP3"},"hgnc":{"alias_symbol":["SGP28","CRISP-3","CRS3","dJ442L6.3","Aeg2"],"prev_symbol":[]},"alphafold":{"accession":"P54108","domains":[{"cath_id":"3.40.33.10","chopping":"27-173","consensus_level":"high","plddt":97.4207,"start":27,"end":173},{"cath_id":"1.10.10.740","chopping":"208-241","consensus_level":"high","plddt":94.6921,"start":208,"end":241}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P54108","model_url":"https://alphafold.ebi.ac.uk/files/AF-P54108-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P54108-F1-predicted_aligned_error_v6.png","plddt_mean":92.5},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CRISP3","jax_strain_url":"https://www.jax.org/strain/search?query=CRISP3"},"sequence":{"accession":"P54108","fasta_url":"https://rest.uniprot.org/uniprotkb/P54108.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P54108/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P54108"}},"corpus_meta":[{"pmid":"8665901","id":"PMC_8665901","title":"The human cysteine-rich secretory protein (CRISP) family. Primary structure and tissue distribution of CRISP-1, CRISP-2 and CRISP-3.","date":"1996","source":"European journal of biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/8665901","citation_count":182,"is_preprint":false},{"pmid":"8319566","id":"PMC_8319566","title":"Transcripts for cysteine-rich secretory protein-1 (CRISP-1; DE/AEG) and the novel related CRISP-3 are expressed under androgen control in the mouse salivary gland.","date":"1993","source":"Endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/8319566","citation_count":110,"is_preprint":false},{"pmid":"8601434","id":"PMC_8601434","title":"SGP28, a novel matrix glycoprotein in specific granules of human neutrophils with similarity to a human testis-specific gene product and a rodent sperm-coating glycoprotein.","date":"1996","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/8601434","citation_count":83,"is_preprint":false},{"pmid":"12009203","id":"PMC_12009203","title":"An ELISA for SGP28/CRISP-3, a cysteine-rich secretory protein in human neutrophils, plasma, and exocrine secretions.","date":"2002","source":"Journal of immunological methods","url":"https://pubmed.ncbi.nlm.nih.gov/12009203","citation_count":73,"is_preprint":false},{"pmid":"12223513","id":"PMC_12223513","title":"Identification of human cysteine-rich secretory protein 3 (CRISP-3) as a matrix protein in a subset of peroxidase-negative granules of neutrophils and in the granules of eosinophils.","date":"2002","source":"Journal of leukocyte biology","url":"https://pubmed.ncbi.nlm.nih.gov/12223513","citation_count":73,"is_preprint":false},{"pmid":"15867000","id":"PMC_15867000","title":"Characterization and localization of cysteine-rich secretory protein 3 (CRISP-3) in the human male reproductive tract.","date":"2005","source":"Journal of andrology","url":"https://pubmed.ncbi.nlm.nih.gov/15867000","citation_count":66,"is_preprint":false},{"pmid":"9748582","id":"PMC_9748582","title":"Equine CRISP-3: primary structure and expression in the male genital tract.","date":"1998","source":"Biochimica et biophysica acta","url":"https://pubmed.ncbi.nlm.nih.gov/9748582","citation_count":60,"is_preprint":false},{"pmid":"15950934","id":"PMC_15950934","title":"beta-Microseminoprotein binds CRISP-3 in human seminal plasma.","date":"2005","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/15950934","citation_count":58,"is_preprint":false},{"pmid":"8887646","id":"PMC_8887646","title":"CRISP-3, a protein with homology to plant defense proteins, is expressed in mouse B cells under the control of Oct2.","date":"1996","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/8887646","citation_count":57,"is_preprint":false},{"pmid":"21389342","id":"PMC_21389342","title":"Equine CRISP3 modulates interaction between spermatozoa and polymorphonuclear neutrophils.","date":"2011","source":"Biology of reproduction","url":"https://pubmed.ncbi.nlm.nih.gov/21389342","citation_count":55,"is_preprint":false},{"pmid":"17433013","id":"PMC_17433013","title":"A polymorphism within the equine CRISP3 gene is associated with stallion fertility in Hanoverian warmblood horses.","date":"2007","source":"Animal genetics","url":"https://pubmed.ncbi.nlm.nih.gov/17433013","citation_count":49,"is_preprint":false},{"pmid":"21814574","id":"PMC_21814574","title":"Cysteine-rich secretory protein-3 (CRISP3) is strongly up-regulated in prostate carcinomas with the TMPRSS2-ERG fusion gene.","date":"2011","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/21814574","citation_count":37,"is_preprint":false},{"pmid":"7639699","id":"PMC_7639699","title":"Isolation and characterization of the androgen-dependent mouse cysteine-rich secretory protein-3 (CRISP-3) gene.","date":"1995","source":"The Biochemical 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diseases","url":"https://pubmed.ncbi.nlm.nih.gov/20680031","citation_count":25,"is_preprint":false},{"pmid":"36263172","id":"PMC_36263172","title":"Circular RNA hsa_circ_0003823 promotes the Tumor Progression, Metastasis and Apatinib Resistance of Esophageal Squamous Cell Carcinoma by miR-607/CRISP3 Axis.","date":"2022","source":"International journal of biological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/36263172","citation_count":24,"is_preprint":false},{"pmid":"25715794","id":"PMC_25715794","title":"Endometrial CRISP3 is regulated throughout the mouse estrous and human menstrual cycle and facilitates adhesion and proliferation of endometrial epithelial cells.","date":"2015","source":"Biology of reproduction","url":"https://pubmed.ncbi.nlm.nih.gov/25715794","citation_count":22,"is_preprint":false},{"pmid":"32357309","id":"PMC_32357309","title":"CRISP3 expression drives prostate cancer invasion and progression.","date":"2020","source":"Endocrine-related cancer","url":"https://pubmed.ncbi.nlm.nih.gov/32357309","citation_count":21,"is_preprint":false},{"pmid":"28855043","id":"PMC_28855043","title":"Association of the cysteine-rich secretory protein-3 (CRISP-3) and some of its polymorphisms with the quality of cryopreserved stallion semen.","date":"2018","source":"Reproduction, fertility, and development","url":"https://pubmed.ncbi.nlm.nih.gov/28855043","citation_count":19,"is_preprint":false},{"pmid":"22740859","id":"PMC_22740859","title":"Copy number changes of CRISP3 in oral squamous cell carcinoma.","date":"2011","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/22740859","citation_count":16,"is_preprint":false},{"pmid":"20676114","id":"PMC_20676114","title":"Growth inhibition mediated by PSP94 or CRISP-3 is prostate cancer cell line specific.","date":"2010","source":"Asian journal of andrology","url":"https://pubmed.ncbi.nlm.nih.gov/20676114","citation_count":15,"is_preprint":false},{"pmid":"33620707","id":"PMC_33620707","title":"MiR-182-5p, MiR-192-5p, and MiR-493-5p Constitute a Regulatory Network with CRISP3 in Seminal Plasma Fluid of Teratozoospermia Patients.","date":"2021","source":"Reproductive sciences (Thousand Oaks, Calif.)","url":"https://pubmed.ncbi.nlm.nih.gov/33620707","citation_count":14,"is_preprint":false},{"pmid":"24573035","id":"PMC_24573035","title":"Eukaryotic expression, purification and structure/function analysis of native, recombinant CRISP3 from human and mouse.","date":"2014","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/24573035","citation_count":12,"is_preprint":false},{"pmid":"20116414","id":"PMC_20116414","title":"Human CRISP-3 binds serum alpha(1)B-glycoprotein across species.","date":"2010","source":"Biochimica et biophysica acta","url":"https://pubmed.ncbi.nlm.nih.gov/20116414","citation_count":11,"is_preprint":false},{"pmid":"19026612","id":"PMC_19026612","title":"A model of the complex between human beta-microseminoprotein and CRISP-3 based on NMR data.","date":"2008","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/19026612","citation_count":11,"is_preprint":false},{"pmid":"29477539","id":"PMC_29477539","title":"Androgen receptor mediated epigenetic regulation of CRISP3 promoter in prostate cancer cells.","date":"2018","source":"The Journal of steroid biochemistry and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/29477539","citation_count":10,"is_preprint":false},{"pmid":"29552153","id":"PMC_29552153","title":"In silico analysis identifies CRISP3 as a potential peripheral blood biomarker for multiple myeloma: From data modeling to validation with RT-PCR.","date":"2018","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/29552153","citation_count":10,"is_preprint":false},{"pmid":"27746003","id":"PMC_27746003","title":"Expression and localization of cysteine-rich secretory protein-3 (CRISP-3) in the prepubertal and postpubertal male horse.","date":"2016","source":"Theriogenology","url":"https://pubmed.ncbi.nlm.nih.gov/27746003","citation_count":10,"is_preprint":false},{"pmid":"27825912","id":"PMC_27825912","title":"Purification and characterization of CRISP-3 from human seminal plasma and its real-time binding kinetics with PSP94.","date":"2016","source":"Journal of chromatography. B, Analytical technologies in the biomedical and life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/27825912","citation_count":10,"is_preprint":false},{"pmid":"27440996","id":"PMC_27440996","title":"Variation in PTCHD2, CRISP3, NAP1L4, FSCB, and AP3B2 associated with spherical equivalent.","date":"2016","source":"Molecular vision","url":"https://pubmed.ncbi.nlm.nih.gov/27440996","citation_count":9,"is_preprint":false},{"pmid":"22993349","id":"PMC_22993349","title":"The expression of β-microseminoprotein but not CRISP3 is reduced in ovarian cancer and correlates to survival.","date":"2012","source":"Anticancer research","url":"https://pubmed.ncbi.nlm.nih.gov/22993349","citation_count":8,"is_preprint":false},{"pmid":"23375721","id":"PMC_23375721","title":"Mapping of the binding sites involved in PSP94-CRISP-3 interaction by molecular dissection of the complex.","date":"2013","source":"Biochimica et biophysica acta","url":"https://pubmed.ncbi.nlm.nih.gov/23375721","citation_count":8,"is_preprint":false},{"pmid":"38396941","id":"PMC_38396941","title":"Semen Protein CRISP3 Promotes Reproductive Performance of Boars through Immunomodulation.","date":"2024","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/38396941","citation_count":6,"is_preprint":false},{"pmid":"31965650","id":"PMC_31965650","title":"Upregulation of CRISP-3 and kallikrein in stallion seminal plasma is associated with poor tolerance of cooled storage.","date":"2020","source":"Reproduction in domestic animals = Zuchthygiene","url":"https://pubmed.ncbi.nlm.nih.gov/31965650","citation_count":6,"is_preprint":false},{"pmid":"39139989","id":"PMC_39139989","title":"Computational insights into CRISP3 downregulation in cervical cancer and its cervical lineages pattern.","date":"2024","source":"Precision clinical medicine","url":"https://pubmed.ncbi.nlm.nih.gov/39139989","citation_count":5,"is_preprint":false},{"pmid":"38377715","id":"PMC_38377715","title":"The role of equine seminal plasma derived cysteine rich secretory protein 3 (CRISP3) in the interaction between polymorphonuclear neutrophils (PMNs) and populations of viable or dead spermatozoa, and bacteria.","date":"2024","source":"Theriogenology","url":"https://pubmed.ncbi.nlm.nih.gov/38377715","citation_count":5,"is_preprint":false},{"pmid":"38879857","id":"PMC_38879857","title":"EP300 promotes tumor stemness via epigenetic activation of CRISP3 leading to lobaplatin resistance in triple-negative breast cancer.","date":"2024","source":"Human cell","url":"https://pubmed.ncbi.nlm.nih.gov/38879857","citation_count":4,"is_preprint":false},{"pmid":"39624089","id":"PMC_39624089","title":"Upregulation of CRISP3 and its clinical values in adult sepsis: a comprehensive analysis based on microarrays and a two-retrospective-cohort study.","date":"2024","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/39624089","citation_count":1,"is_preprint":false},{"pmid":"40473221","id":"PMC_40473221","title":"CRISP3-PSP94 complex regulates P2RX7 mediated signalling in prostate cancer cells and macrophages via CITED2.","date":"2025","source":"Biochimica et biophysica acta. Molecular cell research","url":"https://pubmed.ncbi.nlm.nih.gov/40473221","citation_count":1,"is_preprint":false},{"pmid":"32880450","id":"PMC_32880450","title":"High-Pressure Synthesis and Crystal Structure of the Sulfur-Richest Chromium Sulfide CrS3 Composed of Cr(III) and Disulfide Ions.","date":"2020","source":"Inorganic chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/32880450","citation_count":1,"is_preprint":false},{"pmid":"39407057","id":"PMC_39407057","title":"Characterizing the Impact of Dysregulated Micrornas on CRISP3 Isoforms in Male Infertility.","date":"2024","source":"Reproductive sciences (Thousand Oaks, Calif.)","url":"https://pubmed.ncbi.nlm.nih.gov/39407057","citation_count":0,"is_preprint":false},{"pmid":"41751369","id":"PMC_41751369","title":"CRISP3, a Potential Tumor Suppressor, Inhibits the Progression of High-Grade Serous Ovarian Carcinoma by Modulating the PI3K/AKT Pathway.","date":"2026","source":"Biomedicines","url":"https://pubmed.ncbi.nlm.nih.gov/41751369","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":23069,"output_tokens":5268,"usd":0.074113},"stage2":{"model":"claude-opus-4-6","input_tokens":8856,"output_tokens":3236,"usd":0.18777},"total_usd":0.261883,"stage1_batch_id":"msgbatch_011kfMfE81o6CTcyhVUqumuW","stage2_batch_id":"msgbatch_01DTputS6WNkH745x1nVSoBo","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1996,\n      \"finding\": \"CRISP3 (SGP28) is localized in specific granules of human neutrophils, as determined by subcellular fractionation of human neutrophils and partial primary structure determination of the purified 28 kDa glycoprotein.\",\n      \"method\": \"Protein purification from neutrophil exocytosed material, cDNA cloning, subcellular fractionation\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — direct subcellular fractionation with protein purification and sequencing, replicated in subsequent studies\",\n      \"pmids\": [\"8601434\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"CRISP3 is a matrix protein localized in a subset of peroxidase-negative granules with overlapping characteristics of specific and gelatinase granules in neutrophils, and is also present in granules of eosinophils; it exists as both glycosylated and unglycosylated forms with identical subcellular distribution.\",\n      \"method\": \"Three-layer Percoll density gradient fractionation, secretagogue release studies, double-labeling immunogold electron microscopy\",\n      \"journal\": \"Journal of leukocyte biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — multiple orthogonal methods including immunogold EM and subcellular fractionation with functional mobilization studies\",\n      \"pmids\": [\"12223513\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"Beta-microseminoprotein (MSP/PSP94) and CRISP3 form stable, non-covalent high-affinity complexes in human seminal plasma, with CRISP3 binding MSP through its amino-terminal SCP-domain.\",\n      \"method\": \"Immunoprecipitation, gel filtration, surface plasmon resonance\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — reciprocal Co-IP, gel filtration, and quantitative SPR binding kinetics in a single study\",\n      \"pmids\": [\"15950934\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1996,\n      \"finding\": \"CRISP3 expression in mouse B cells is transcriptionally activated by the Oct2 transcription factor via two variant octamer motifs in the CRISP3 promoter; the C-terminal transactivation domain of Oct2 is required, and CRISP3 is specifically expressed at the pre-B-cell stage.\",\n      \"method\": \"Nuclear run-on experiments, Oct2-deficient primary B cells, cotransfection reporter assays, site-directed mutagenesis of octamer motifs, in vitro Oct2–DNA binding\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — multiple orthogonal methods including nuclear run-on, knockout primary cells, and mutagenesis of regulatory elements\",\n      \"pmids\": [\"8887646\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1993,\n      \"finding\": \"CRISP3 gene expression in mouse salivary gland is strongly androgen-dependent, as demonstrated by castration abolishing/reducing expression in male mice and absence of expression in female salivary gland.\",\n      \"method\": \"cDNA cloning, RNA blot analysis of castrated vs. intact males and females\",\n      \"journal\": \"Endocrinology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct hormonal manipulation with quantitative RNA blotting, single lab\",\n      \"pmids\": [\"8319566\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1995,\n      \"finding\": \"The mouse CRISP3 gene spans over 20 kb, consists of eight exons and seven introns, has a TATA box-proximal transcription start site, and contains two androgen-responsive element consensus sequences in its promoter along with putative OTF- and GATA-binding elements.\",\n      \"method\": \"Genomic library screening, DNA sequencing, primer extension mapping, PCR sizing of introns\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct genomic characterization with promoter element identification\",\n      \"pmids\": [\"7639699\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"NMR analysis of the MSP–CRISP3 complex shows that only one side of MSP (comprising beta-strands 1, 4, 5, and 8, with beta-strands 1 and 8 forming the main binding surface) is affected by complex formation with the N-terminal SCP domain of CRISP3.\",\n      \"method\": \"Multidimensional NMR (15N-HSQC and 3D-NMR of triply-labeled MSP in complex with recombinant N-terminal CRISP3 domain)\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 — NMR structural characterization of the complex, but model is tentative (no full CRISP3 structure)\",\n      \"pmids\": [\"19026612\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Equine CRISP3 purified from seminal plasma suppresses binding between spermatozoa and polymorphonuclear neutrophils (PMNs), identifying CRISP3 as a seminal plasma protein that modulates sperm elimination from the female reproductive tract.\",\n      \"method\": \"Sequential ammonium sulfate precipitation, size-exclusion and ion-exchange chromatography, 1D/2D SDS-PAGE, Western blotting, flow cytometry PMN/sperm binding assay\",\n      \"journal\": \"Biology of reproduction\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — protein purified to homogeneity, functional activity confirmed by flow cytometry with appropriate controls\",\n      \"pmids\": [\"21389342\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Human CRISP3 binds alpha-1B-glycoprotein (A1BG) in serum; A1BG orthologs from cow, horse, and rabbit also bind CRISP3, while mouse kininogen-1 binds CRISP3 in mouse serum, suggesting a conserved mechanism of CRISP binding by A1BG-family proteins.\",\n      \"method\": \"Gel filtration, ligand blotting, affinity isolation, mass spectrometry, N-terminal sequencing\",\n      \"journal\": \"Biochimica et biophysica acta\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — affinity isolation with MS identification across multiple species, single lab\",\n      \"pmids\": [\"20116414\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"The binding interface of PSP94–CRISP3 complex was mapped: PSP94 residues Y3, F4, P56, and its C-terminal beta-strand (plus the C37–C73 disulfide bond) are critical for CRISP3 binding; on CRISP3, the N-terminal SCP domain alone is insufficient—the hinge region is also required, but the C-terminal ICR domain is not.\",\n      \"method\": \"Site-directed mutagenesis of PSP94 and CRISP3, co-immunoprecipitation\",\n      \"journal\": \"Biochimica et biophysica acta\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — mutagenesis combined with Co-IP to map binding residues, single lab\",\n      \"pmids\": [\"23375721\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Native, glycosylated human and mouse CRISP3 expressed in HEK 293 cells exist as monomers in solution; N-glycosylation sites and patterns differ between human and mouse CRISP3.\",\n      \"method\": \"Eukaryotic expression in HEK 293 cells, ion exchange and size exclusion chromatography, substrate-affinity assays, glycosylation characterization\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — native protein purification with functional validation, single lab\",\n      \"pmids\": [\"24573035\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"CRISP3 is a direct transcriptional target of the ERG transcription factor (driven by TMPRSS2-ERG fusion) in prostate cancer, as shown by chromatin immunoprecipitation with an anti-ERG antibody demonstrating ERG binding at the CRISP3 locus.\",\n      \"method\": \"Genome-wide mRNA expression analysis, quantitative RT-PCR, immunohistochemistry, chromatin immunoprecipitation (ChIP) with anti-ERG antibody\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP directly demonstrates ERG occupancy at CRISP3 locus, supported by expression correlation\",\n      \"pmids\": [\"21814574\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"CRISP3 expression in prostate cancer cells is androgen-dependent and regulated by androgen receptor (AR) through epigenetic mechanisms: in AR-negative cells the CRISP3 promoter is silenced by histone deacetylation; DHT treatment of LNCaP cells increases CRISP3 transcript and protein; AR occupancy at the CRISP3 promoter was confirmed by ChIP.\",\n      \"method\": \"Luciferase reporter assays with CRISP3 promoter fragments, ChIP-PCR, DHT treatment, histone deacetylase inhibition\",\n      \"journal\": \"The Journal of steroid biochemistry and molecular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (reporter assay, ChIP, hormone treatment) in a single lab\",\n      \"pmids\": [\"29477539\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Recombinant CRISP3 enhances adhesion and proliferation of human endometrial epithelial cells in vitro; CRISP3 is secreted by primary human endometrial epithelial cells and accumulates in uterine lavage fluid, with higher abundance during the proliferative phase.\",\n      \"method\": \"Treatment of endometrial epithelial cells with recombinant CRISP3, adhesion and proliferation assays, uterine lavage ELISA, immunohistochemistry\",\n      \"journal\": \"Biology of reproduction\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — recombinant protein functional assay with defined cellular readouts, single lab\",\n      \"pmids\": [\"25715794\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"CRISP3 facilitates prostate cancer progression from carcinoma in situ to invasive cancer in vivo; CRISP3 exposure enhances cancer cell motility and invasion; mass spectrometry revealed CRISP3 induces changes in abundance of cell-cell adhesion proteins LASP1 and TJP1 both in vivo and in vitro.\",\n      \"method\": \"Genetically engineered mouse model of prostate cancer, human and mouse prostate cancer cell motility/invasion assays, mass spectrometry proteomics\",\n      \"journal\": \"Endocrine-related cancer\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo mouse model combined with cell-based functional assays and proteomics, multiple orthogonal approaches\",\n      \"pmids\": [\"32357309\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"PSP94 and CRISP3 each independently inhibit prostate cancer cell growth in a cell-line-specific manner; growth inhibition by CRISP3 is not affected by presence or absence of PSP94, suggesting CRISP3 participates in PSP94-independent activities.\",\n      \"method\": \"Ectopic expression of PSP94 and CRISP3 in prostate cell lines, clonogenic survival assay, Western blot, immunofluorescence\",\n      \"journal\": \"Asian journal of andrology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — gain-of-function with defined cellular phenotype, single lab\",\n      \"pmids\": [\"20676114\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"CRISP3 was purified from human seminal plasma using PSP94-immobilized affinity chromatography and confirmed to bind PSP94 with high affinity by surface plasmon resonance; it exists as glycosylated (~28 kDa) and unglycosylated (~26 kDa) forms.\",\n      \"method\": \"PSP94-affinity chromatography, SDS-PAGE, immunoblotting, MALDI-TOF MS, deglycosylation, surface plasmon resonance\",\n      \"journal\": \"Journal of chromatography B\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — protein purified to homogeneity with binding kinetics validated by SPR\",\n      \"pmids\": [\"27825912\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Exogenous CRISP3 downregulates P2RX7 (an ATP-gated ion channel) in PC3 prostate cancer cells and THP1 macrophages, reducing ATP-induced cytotoxicity and IL-1β secretion; this effect is abrogated when CRISP3 is complexed with PSP94. CRISP3 mediates these effects through CITED2 (a transcriptional coregulator that reduces p300 availability at the P2RX7 promoter); PSP94 also affects CRISP3 endocytosis and its interaction with flotillin-2.\",\n      \"method\": \"Recombinant protein treatment, P2RX7 expression assays, ATP cytotoxicity assay, IL-1β ELISA, antibody array, CITED2 overexpression, CUT&RUN assay for p300 at P2RX7 promoter, Co-IP with flotillin-2\",\n      \"journal\": \"Biochimica et biophysica acta. Molecular cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal mechanistic methods in a single lab defining a novel signaling axis\",\n      \"pmids\": [\"40473221\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Boar CRISP3 is localized in the post-acrosomal region of the sperm head and migrates to the anterior end of the tail after capacitation; recombinant CRISP3 downregulates inflammatory factors IL-1α, IL-1β, and IL-6 in LPS-stimulated RAW264.7 macrophages, indicating immunomodulatory function.\",\n      \"method\": \"Immunofluorescence localization, recombinant protein treatment of macrophages, RT-PCR and Western blot for inflammatory cytokines\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct localization experiment linked to functional immunomodulatory readout, single lab\",\n      \"pmids\": [\"38396941\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Equine seminal plasma CRISP3 selectively suppresses PMN binding to live spermatozoa but not to dead spermatozoa or bacteria; immunocytochemistry confirmed CRISP3 binds to live but not dead spermatozoa, suggesting selective recognition of viable sperm surface.\",\n      \"method\": \"CRISP3 purification from seminal plasma, flow cytometry PMN binding assay with live/dead sperm populations, immunocytochemistry\",\n      \"journal\": \"Theriogenology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — purified protein functional assay with flow cytometry and localization, single lab\",\n      \"pmids\": [\"38377715\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"EP300 histone acetyltransferase regulates CRISP3 expression in triple-negative breast cancer by modifying H3K27ac at the CRISP3 promoter; CRISP3 overexpression promotes tumor stemness and lobaplatin resistance, and knockdown of EP300 reduces these phenotypes in a manner rescued by CRISP3 overexpression.\",\n      \"method\": \"ChIP for H3K27ac at CRISP3 promoter, EP300 knockdown and CRISP3 overexpression rescue experiments, in vitro and in vivo stemness/resistance assays\",\n      \"journal\": \"Human cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP-based epigenetic mechanism with epistatic rescue experiment, single lab\",\n      \"pmids\": [\"38879857\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"hsa_circ_0003823 promotes ESCC progression and apatinib resistance through the miR-607/CRISP3 axis: the circular RNA sequesters miR-607, leading to upregulation of CRISP3; CRISP3 was validated as a miR-607 target by RNA immunoprecipitation and dual-luciferase reporter assays.\",\n      \"method\": \"RNA immunoprecipitation (RIP), dual-luciferase reporter assay, Western blot, knockdown experiments\",\n      \"journal\": \"International journal of biological sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — RIP and luciferase assays directly demonstrate miR-607 targeting of CRISP3, single lab\",\n      \"pmids\": [\"36263172\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"CRISP3 acts as a tumor suppressor in high-grade serous ovarian carcinoma by inhibiting cell proliferation, migration, and invasion through suppression of the PI3K/AKT signaling pathway; CRISP3 knockdown activates PI3K/AKT and overexpression suppresses it.\",\n      \"method\": \"CRISP3 knockdown and overexpression in HGSOC cells, CCK-8/EdU proliferation and Transwell invasion assays, RNA-seq, GSEA, Western blotting for PI3K/AKT pathway components, in vivo mouse model\",\n      \"journal\": \"Biomedicines\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — loss- and gain-of-function with pathway readout by Western blot and RNA-seq, single lab\",\n      \"pmids\": [\"41751369\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CRISP3 is a secreted, cysteine-rich glycoprotein (with glycosylated and unglycosylated forms) that is stored in neutrophil specific/gelatinase granules and exocrine secretions; its expression is controlled by androgens (via AR-mediated epigenetic regulation), Oct2, ERG (in TMPRSS2-ERG fusion prostate cancer), and EP300-mediated H3K27 acetylation; it forms high-affinity non-covalent complexes with MSP/PSP94 through its N-terminal SCP domain and hinge region, binds serum A1BG, and functionally modulates immune responses by suppressing PMN–sperm binding, downregulating inflammatory cytokines, and reducing P2RX7-mediated IL-1β secretion through a CITED2/p300 axis; it also influences cell adhesion (via LASP1, TJP1), promotes prostate cancer invasion, and suppresses ovarian cancer progression via the PI3K/AKT pathway.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"CRISP3 is a secreted cysteine-rich glycoprotein that functions at the intersection of innate immunity, reproductive biology, and cancer progression. It is stored in specific and gelatinase granules of neutrophils and eosinophils, from which it is released upon activation, and is also secreted by exocrine glands and endometrial epithelium [PMID:8601434, PMID:12223513, PMID:25715794]. CRISP3 forms high-affinity non-covalent complexes with PSP94/MSP through its N-terminal SCP domain and hinge region, and binds the serum carrier alpha-1B-glycoprotein (A1BG); the PSP94 interaction modulates CRISP3 activity, including its ability to downregulate P2RX7 expression via a CITED2/p300 transcriptional axis, thereby suppressing IL-1β secretion [PMID:15950934, PMID:23375721, PMID:20116414, PMID:40473221]. Its expression is controlled by androgens via AR-mediated promoter occupancy, by the ERG transcription factor in TMPRSS2-ERG fusion-positive prostate cancers, and by EP300-dependent H3K27 acetylation; functionally, CRISP3 suppresses PMN–sperm binding to protect viable spermatozoa, downregulates inflammatory cytokines in macrophages, promotes prostate cancer invasion by altering cell-adhesion proteins LASP1 and TJP1, and suppresses ovarian cancer progression through inhibition of PI3K/AKT signaling [PMID:29477539, PMID:21814574, PMID:38879857, PMID:21389342, PMID:38396941, PMID:32357309, PMID:41751369].\",\n  \"teleology\": [\n    {\n      \"year\": 1993,\n      \"claim\": \"Establishing that CRISP3 is an androgen-regulated gene answered the question of what controls its tissue-specific expression in exocrine glands.\",\n      \"evidence\": \"Castration and RNA blot analysis in mouse salivary gland\",\n      \"pmids\": [\"8319566\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single tissue (salivary gland) examined\", \"Androgen-responsive elements in promoter not functionally validated\"]\n    },\n    {\n      \"year\": 1996,\n      \"claim\": \"Identification of CRISP3 as a specific granule protein of neutrophils established it as a component of the innate immune arsenal, while independent work showed Oct2-driven transcription in pre-B cells revealed a second lineage of expression.\",\n      \"evidence\": \"Subcellular fractionation and protein purification from neutrophils; nuclear run-on, Oct2-knockout B cells, and reporter assays in mouse\",\n      \"pmids\": [\"8601434\", \"8887646\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Function of CRISP3 after neutrophil degranulation unknown\", \"Whether Oct2-driven B-cell expression is conserved in humans\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Refined granule subtyping showed CRISP3 resides in a subset of peroxidase-negative granules overlapping specific and gelatinase granules, and extended expression to eosinophils, broadening its potential role in granulocyte biology.\",\n      \"evidence\": \"Three-layer Percoll fractionation, secretagogue mobilization, double-labeling immunogold EM in human neutrophils and eosinophils\",\n      \"pmids\": [\"12223513\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of CRISP3 release from granulocyte granules not tested\", \"Receptor or target on effector cells unknown\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Discovery that CRISP3 forms a stable high-affinity complex with PSP94/MSP in seminal plasma via its SCP domain identified a major extracellular binding partner and suggested functional modulation of both proteins.\",\n      \"evidence\": \"Reciprocal co-immunoprecipitation, gel filtration, and surface plasmon resonance in human seminal plasma\",\n      \"pmids\": [\"15950934\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Biological consequence of complex formation unknown\", \"Binding stoichiometry in vivo not established\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"NMR mapping of the PSP94–CRISP3 interface revealed which PSP94 surfaces are engaged, providing the first structural picture of CRISP3 complex formation.\",\n      \"evidence\": \"Multidimensional NMR of 15N/13C/2H-labeled PSP94 in complex with recombinant CRISP3 N-terminal domain\",\n      \"pmids\": [\"19026612\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Full CRISP3 structure not determined\", \"NMR model tentative without full-length complex structure\", \"Hinge region contribution not captured\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Two advances clarified CRISP3's broader interaction network and its cancer-cell-autonomous activity: identification of A1BG as a serum carrier across species, and demonstration that CRISP3 can independently inhibit prostate cancer cell growth.\",\n      \"evidence\": \"Gel filtration/affinity isolation/MS for A1BG binding; ectopic expression and clonogenic assays in prostate cancer cells\",\n      \"pmids\": [\"20116414\", \"20676114\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of A1BG–CRISP3 interaction unknown\", \"Mechanism of growth inhibition undefined\", \"Both findings from single laboratories\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Two studies established CRISP3's roles in reproduction and oncogenic transcription: equine CRISP3 suppresses PMN–sperm binding, protecting spermatozoa, while ERG directly transactivates CRISP3 in TMPRSS2-ERG fusion prostate cancer.\",\n      \"evidence\": \"Purified protein flow cytometry PMN/sperm binding assay; ChIP with anti-ERG antibody at CRISP3 locus in prostate cancer\",\n      \"pmids\": [\"21389342\", \"21814574\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"CRISP3 receptor on PMNs or sperm not identified\", \"Whether ERG-driven CRISP3 is functionally required for prostate cancer phenotype not tested\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Detailed mutagenesis of both PSP94 and CRISP3 established that the SCP domain plus hinge region of CRISP3 (but not ICR) and specific PSP94 residues are critical for complex formation, refining the structural model.\",\n      \"evidence\": \"Site-directed mutagenesis combined with co-immunoprecipitation\",\n      \"pmids\": [\"23375721\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No crystal structure of the complex\", \"Functional significance of each binding-site residue untested in vivo\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"AR-mediated epigenetic regulation of CRISP3 in prostate cancer was demonstrated, showing AR occupancy at the CRISP3 promoter and that histone deacetylation silences CRISP3 in AR-negative cells.\",\n      \"evidence\": \"ChIP-PCR for AR, luciferase reporters, DHT treatment and HDAC inhibition in LNCaP cells\",\n      \"pmids\": [\"29477539\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Specific histone marks at the CRISP3 promoter not mapped in this study\", \"Relationship between AR and ERG-driven regulation not addressed\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"An in vivo prostate cancer model demonstrated that CRISP3 promotes progression from carcinoma in situ to invasive cancer, mechanistically linked to altered abundance of cell-adhesion proteins LASP1 and TJP1.\",\n      \"evidence\": \"Genetically engineered mouse model, cell motility/invasion assays, mass spectrometry proteomics\",\n      \"pmids\": [\"32357309\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct interaction of CRISP3 with LASP1 or TJP1 not shown\", \"Whether CRISP3 acts intracellularly or extracellularly to modulate adhesion proteins unclear\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Multiple 2024 studies collectively expanded the functional repertoire: CRISP3 downregulates P2RX7 via CITED2/p300 to suppress IL-1β secretion (blocked when complexed with PSP94); boar CRISP3 downregulates inflammatory cytokines in macrophages; equine CRISP3 selectively protects live sperm from PMN attack; and EP300-mediated H3K27ac drives CRISP3 expression that promotes stemness and drug resistance in triple-negative breast cancer.\",\n      \"evidence\": \"CUT&RUN for p300 at P2RX7, Co-IP with flotillin-2, macrophage cytokine assays, flow cytometry with live/dead sperm, ChIP for H3K27ac at CRISP3 promoter with epistatic rescue in TNBC\",\n      \"pmids\": [\"40473221\", \"38396941\", \"38377715\", \"38879857\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"CITED2/p300 axis validated in one cell system only\", \"Surface receptor mediating CRISP3 signaling into target cells unidentified\", \"EP300/CRISP3 axis in breast cancer not independently replicated\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"CRISP3 was shown to function as a tumor suppressor in high-grade serous ovarian carcinoma by inhibiting PI3K/AKT signaling, contrasting with its pro-invasive role in prostate cancer and revealing context-dependent oncogenic behavior.\",\n      \"evidence\": \"Loss- and gain-of-function in HGSOC cells, RNA-seq/GSEA, PI3K/AKT pathway Western blotting, in vivo mouse xenograft\",\n      \"pmids\": [\"41751369\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab finding not independently replicated\", \"Mechanism by which CRISP3 suppresses PI3K/AKT not defined\", \"Contradiction with pro-tumorigenic roles in other cancers unexplained\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The cell-surface receptor(s) through which extracellular CRISP3 signals remain unidentified, and there is no high-resolution structure of full-length CRISP3 or its complexes.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No receptor identified on neutrophils, sperm, or cancer cells\", \"No crystal or cryo-EM structure of full-length CRISP3\", \"Context-dependent tumor-promoting versus tumor-suppressing activities mechanistically unexplained\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [7, 17, 19, 22]},\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [13, 14, 17, 18]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [2, 7, 8, 16, 19]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": []},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [0, 1, 7, 17, 18, 19]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [17, 22]},\n      {\"term_id\": \"R-HSA-1474165\", \"supporting_discovery_ids\": [7, 19]}\n    ],\n    \"complexes\": [\n      \"PSP94-CRISP3 complex\",\n      \"A1BG-CRISP3 complex\"\n    ],\n    \"partners\": [\n      \"MSMB\",\n      \"A1BG\",\n      \"CITED2\",\n      \"EP300\",\n      \"FLOT2\",\n      \"LASP1\",\n      \"TJP1\",\n      \"P2RX7\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}