{"gene":"EFCAB7","run_date":"2026-06-09T23:54:42","timeline":{"discoveries":[{"year":2014,"finding":"EFCAB7 forms a complex with IQCE that anchors the EVC-EVC2 heterodimer in a signaling microdomain at the base of primary cilia (the EvC zone). EFCAB7 directly binds a C-terminal disordered region of EVC2 that is deleted in Weyers syndrome patients. Depletion of EFCAB7 causes mislocalization of EVC-EVC2 within cilia and impairs activation of the transcription factor GLI2, phenocopying the Weyers cellular phenotype.","method":"Co-immunoprecipitation, ciliary localization by immunofluorescence, siRNA knockdown with GLI2 activation assay, evolutionary/domain analysis","journal":"Developmental Cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, direct localization experiments, loss-of-function with defined pathway phenotype (GLI2 activation), binding domain mapping; foundational paper with 77 citations and multiple orthogonal methods","pmids":["24582806"],"is_preprint":false},{"year":2023,"finding":"The EvC zone targeting of EVC-EVC2 depends on two separate EFCAB7-binding motifs within EVC2's Weyers-deleted peptide; only one had been previously characterized. SUMOylation of EVC-EVC2 cytosolic tails with SUMO3 enhances complex accumulation at the EvC zone, possibly via increased binding to the EFCAB7-IQCE complex. EFCAB7 and IQCE were confirmed as EVC interactors by endogenous EVC protein interactome profiling in control and Evc-null cells.","method":"Endogenous Co-IP/mass spectrometry (interactome screen in Evc-null vs. control cells), SUMOylation assays, ciliary localization by immunofluorescence, mapping of EVC2 binding motifs","journal":"Frontiers in Cell and Developmental Biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — endogenous interactome confirmed by MS and Co-IP, SUMOylation functional assay, motif mapping; single lab, multiple orthogonal methods","pmids":["37576597"],"is_preprint":false},{"year":2025,"finding":"A splicing variant (c.683-1G>C) in EFCAB7 causes exon skipping and production of a non-functional protein. Mice carrying this variant exhibit impaired ciliogenesis, disrupted Hedgehog/Shh-Gli pathway activity, and transcriptional downregulation of Gli target genes (Myh6, Zfpm1, Nkx2-5), resulting in abnormal cardiac development including Tetralogy of Fallot-like defects.","method":"Sanger sequencing of patients, in vitro and in vivo splicing assays, mouse model with variant knock-in, ciliogenesis assay, CUT&Tag for Gli binding, bulk RNA-seq of embryonic hearts","journal":"The Journal of Biological Chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo mouse model with multiple readouts (ciliogenesis, pathway activity, transcriptomics), single lab, multiple orthogonal methods","pmids":["39894222"],"is_preprint":false},{"year":2024,"finding":"Full deletion of Efcab7 in mice sensitizes fetuses to craniofacial and ocular malformations induced by the Smo antagonist vismodegib or by alcohol, while reducing malformations induced by the Smo agonist SAG. This establishes EFCAB7 as a positive regulator of Smoothened (Smo) activity within the Shh pathway in vivo.","method":"CRISPR/genetic knockout mouse model, pharmacological challenge with Smo agonist/antagonist and alcohol at gastrulation stage, fetal phenotypic scoring","journal":"Reproductive Toxicology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean KO mouse with defined epistatic relationship to Smo agonist/antagonist, single lab, genetic and pharmacological dissection","pmids":["39366525"],"is_preprint":false},{"year":2024,"finding":"EFCAB7 directly interacts with PARK7 (DJ-1) in hepatocellular carcinoma cells, as demonstrated by mass spectrometry and Co-IP. Overexpression of PARK7 in EFCAB7-knockdown cells rescues tumor cell proliferation and metastasis, indicating EFCAB7 acts upstream of PARK7 in a cancer cell context.","method":"Mass spectrometry, Co-immunoprecipitation, siRNA knockdown, PARK7 rescue overexpression, in vitro proliferation/metastasis assays","journal":"Aging","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single Co-IP and MS from one lab, no structural or reconstitution validation, context is cancer cell line only","pmids":["39276379"],"is_preprint":false},{"year":2025,"finding":"CRISPR/Cas9-mediated homozygous deletion of Efcab7 in mice produces no detectable defects in male fertility, sperm motility, or testis/epididymis morphology, indicating Efcab7 is dispensable for male reproduction when ablated alone.","method":"CRISPR/Cas9 knockout mice, mating tests, sperm motility analysis, histological phenotyping","journal":"Journal of Biomedical Research","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — clean KO with multiple phenotypic readouts confirming null result; single lab, single study, but rigorous in vivo approach","pmids":["41194443"],"is_preprint":false}],"current_model":"EFCAB7 is a ciliary protein that forms a complex with IQCE to anchor the EVC-EVC2 heterodimer at the EvC zone (base of primary cilia), thereby positively regulating Hedgehog/Smoothened-GLI2 signaling; its loss mislocalizes EVC-EVC2, impairs GLI2 activation, and causes developmental defects including skeletal anomalies, polydactyly, and congenital heart defects, while SUMOylation of EVC-EVC2 and two distinct EFCAB7-binding motifs on EVC2 further modulate complex stability and ciliary targeting."},"narrative":{"mechanistic_narrative":"EFCAB7 is a primary cilium protein that functions as a positive regulator of Hedgehog signaling by anchoring the EVC-EVC2 heterodimer within a specialized signaling microdomain at the ciliary base (the EvC zone) [PMID:24582806]. Together with IQCE, EFCAB7 forms a complex that directly binds a C-terminal disordered region of EVC2 — the same region deleted in Weyers syndrome — and its depletion mislocalizes EVC-EVC2 within cilia and impairs activation of the GLI2 transcription factor [PMID:24582806]. EvC zone targeting depends on two distinct EFCAB7-binding motifs in the EVC2 Weyers-deleted peptide, and SUMOylation of the EVC-EVC2 cytosolic tails enhances complex accumulation at this site, consistent with modulation of binding to the EFCAB7-IQCE complex [PMID:37576597]. In vivo, EFCAB7 acts upstream as a positive regulator of Smoothened activity: full deletion in mice sensitizes fetuses to malformations induced by Smo antagonism and protects against Smo agonism [PMID:39366525], and a disease-associated splicing variant disrupts ciliogenesis, downregulates Gli target genes (Myh6, Zfpm1, Nkx2-5), and produces Tetralogy of Fallot-like cardiac defects [PMID:39894222]. EFCAB7 loss is nonetheless dispensable for male fertility and sperm motility when ablated alone [PMID:41194443].","teleology":[{"year":2014,"claim":"Established how EVC-EVC2 is retained at the ciliary base and linked EFCAB7 to a defined Hedgehog signaling output, answering how a Weyers-syndrome–associated EVC2 region is functionally read out.","evidence":"Co-IP, ciliary immunofluorescence, EFCAB7 siRNA knockdown with GLI2 activation assay, and binding-domain mapping in mammalian cells","pmids":["24582806"],"confidence":"High","gaps":["Does not resolve the structural basis of the EFCAB7-IQCE-EVC2 complex","Stoichiometry and assembly order at the EvC zone not defined"]},{"year":2023,"claim":"Refined the molecular interface and regulation, showing EvC zone targeting requires two separate EFCAB7-binding motifs and is enhanced by SUMOylation of the EVC-EVC2 tails.","evidence":"Endogenous EVC interactome by Co-IP/MS in Evc-null vs control cells, SUMOylation assays, ciliary localization, and EVC2 motif mapping (single lab)","pmids":["37576597"],"confidence":"Medium","gaps":["Mechanism by which SUMO3 increases EFCAB7-IQCE binding not directly demonstrated","SUMO ligase/site responsible not identified","Single lab"]},{"year":2024,"claim":"Placed EFCAB7 epistatically within the Shh pathway in vivo as a positive regulator of Smoothened activity, distinguishing its role from upstream/downstream nodes.","evidence":"CRISPR Efcab7-knockout mice challenged with Smo agonist (SAG), Smo antagonist (vismodegib), or alcohol at gastrulation, with fetal phenotypic scoring","pmids":["39366525"],"confidence":"Medium","gaps":["Does not define the molecular step at which EFCAB7 modulates Smo","Single lab","Epistasis inferred pharmacologically rather than biochemically"]},{"year":2025,"claim":"Connected an EFCAB7 loss-of-function variant to a human-relevant developmental disease mechanism, tying ciliogenesis and Gli-target transcription to cardiac morphogenesis.","evidence":"Splicing assays, variant knock-in mouse, ciliogenesis assay, CUT&Tag for Gli binding, and bulk RNA-seq of embryonic hearts","pmids":["39894222"],"confidence":"Medium","gaps":["Direct effect of EFCAB7 on ciliogenesis machinery not mechanistically resolved","Single lab","Causality between Gli-target downregulation and specific cardiac defects not fully dissected"]},{"year":2024,"claim":"Reported a non-ciliary context in which EFCAB7 interacts with PARK7 and acts upstream of it to support tumor cell proliferation and metastasis.","evidence":"Mass spectrometry and Co-IP in hepatocellular carcinoma cells with siRNA knockdown and PARK7 rescue overexpression (in vitro)","pmids":["39276379"],"confidence":"Low","gaps":["Single Co-IP and MS from one lab, no reciprocal or structural validation","Restricted to one cancer cell line context","Relationship to ciliary function unknown"]},{"year":2025,"claim":"Defined the boundary of EFCAB7 requirement in vivo, showing it is dispensable for male reproduction when ablated alone.","evidence":"CRISPR/Cas9 knockout mice with mating tests, sperm motility analysis, and histological phenotyping","pmids":["41194443"],"confidence":"Medium","gaps":["Possible redundancy with paralogs not tested","Female reproductive phenotype not addressed"]},{"year":null,"claim":"The structural architecture of the EFCAB7-IQCE-EVC2 assembly and the precise molecular step at which EFCAB7 enables Smoothened-to-GLI2 signal transduction remain undefined.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structure of the EFCAB7-IQCE-EVC2 complex","Direct biochemical link between complex anchoring and Smo activation unresolved","PARK7 interaction not integrated with ciliary role"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,1]}],"localization":[{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[0,1]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,2,3]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[2,3]}],"complexes":["EFCAB7-IQCE complex","EvC zone (EVC-EVC2-EFCAB7-IQCE)"],"partners":["IQCE","EVC2","EVC","PARK7"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"A8K855","full_name":"EF-hand calcium-binding domain-containing protein 7","aliases":[],"length_aa":629,"mass_kda":72.0,"function":"Component of the EvC complex that positively regulates ciliary Hedgehog (Hh) signaling. Required for the localization of the EVC2:EVC subcomplex at the base of primary cilia","subcellular_location":"Cell projection, cilium membrane","url":"https://www.uniprot.org/uniprotkb/A8K855/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/EFCAB7","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/EFCAB7","total_profiled":1310},"omim":[{"mim_id":"617632","title":"EF-HAND CALCIUM-BINDING DOMAIN-CONTAINING PROTEIN 7; EFCAB7","url":"https://www.omim.org/entry/617632"},{"mim_id":"617631","title":"IQ DOMAIN-CONTAINING PROTEIN E; IQCE","url":"https://www.omim.org/entry/617631"},{"mim_id":"607261","title":"EVC CILIARY COMPLEX SUBUNIT 2; EVC2","url":"https://www.omim.org/entry/607261"},{"mim_id":"604831","title":"EVC CILIARY COMPLEX SUBUNIT 1; EVC","url":"https://www.omim.org/entry/604831"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Primary cilium","reliability":"Approved"},{"location":"Cytosol","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/EFCAB7"},"hgnc":{"alias_symbol":["KIAA1799","RP4-534K7.1"],"prev_symbol":[]},"alphafold":{"accession":"A8K855","domains":[{"cath_id":"1.10.238,1.10.238","chopping":"32-99","consensus_level":"medium","plddt":87.1854,"start":32,"end":99},{"cath_id":"1.10.238.10","chopping":"105-169","consensus_level":"medium","plddt":88.0805,"start":105,"end":169},{"cath_id":"2.60.40.1180","chopping":"245-258_527-629","consensus_level":"high","plddt":88.9703,"start":245,"end":629},{"cath_id":"2.60.120","chopping":"260-379_507-525","consensus_level":"medium","plddt":89.4173,"start":260,"end":525},{"cath_id":"1.10.238.10","chopping":"384-499","consensus_level":"medium","plddt":93.009,"start":384,"end":499}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/A8K855","model_url":"https://alphafold.ebi.ac.uk/files/AF-A8K855-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-A8K855-F1-predicted_aligned_error_v6.png","plddt_mean":80.25},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=EFCAB7","jax_strain_url":"https://www.jax.org/strain/search?query=EFCAB7"},"sequence":{"accession":"A8K855","fasta_url":"https://rest.uniprot.org/uniprotkb/A8K855.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/A8K855/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/A8K855"}},"corpus_meta":[{"pmid":"24582806","id":"PMC_24582806","title":"EFCAB7 and IQCE regulate hedgehog signaling by tethering the EVC-EVC2 complex to the base of primary cilia.","date":"2014","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/24582806","citation_count":77,"is_preprint":false},{"pmid":"25937073","id":"PMC_25937073","title":"Global analysis of chromosome 1 genes among patients with lung adenocarcinoma, squamous carcinoma, large-cell carcinoma, small-cell carcinoma, or non-cancer.","date":"2015","source":"Cancer metastasis reviews","url":"https://pubmed.ncbi.nlm.nih.gov/25937073","citation_count":59,"is_preprint":false},{"pmid":"26748586","id":"PMC_26748586","title":"Truncation and microdeletion of EVC/EVC2 with missense mutation of EFCAB7 in Ellis-van Creveld syndrome.","date":"2016","source":"Congenital anomalies","url":"https://pubmed.ncbi.nlm.nih.gov/26748586","citation_count":12,"is_preprint":false},{"pmid":"37684519","id":"PMC_37684519","title":"Variants in EFCAB7 underlie nonsyndromic postaxial polydactyly.","date":"2023","source":"European journal of human genetics : EJHG","url":"https://pubmed.ncbi.nlm.nih.gov/37684519","citation_count":8,"is_preprint":false},{"pmid":"22499347","id":"PMC_22499347","title":"Genome-wide linkage and copy number variation analysis reveals 710 kb duplication on chromosome 1p31.3 responsible for autosomal dominant omphalocele.","date":"2012","source":"Journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/22499347","citation_count":8,"is_preprint":false},{"pmid":"37576597","id":"PMC_37576597","title":"EVC-EVC2 complex stability and ciliary targeting are regulated by modification with ubiquitin and SUMO.","date":"2023","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/37576597","citation_count":7,"is_preprint":false},{"pmid":"39894222","id":"PMC_39894222","title":"A splicing variant in EFCAB7 hinders ciliary transport and disrupts cardiac development.","date":"2025","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/39894222","citation_count":2,"is_preprint":false},{"pmid":"39366525","id":"PMC_39366525","title":"Efcab7 deletion sensitizes mice to the teratogenic effects of gastrulation-stage alcohol exposure.","date":"2024","source":"Reproductive toxicology (Elmsford, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/39366525","citation_count":2,"is_preprint":false},{"pmid":"39276379","id":"PMC_39276379","title":"Upregulation of EFCAB7 after radiofrequency ablation promoting hepatocellular carcinoma metastasis and survival by regulating PARK7.","date":"2024","source":"Aging","url":"https://pubmed.ncbi.nlm.nih.gov/39276379","citation_count":0,"is_preprint":false},{"pmid":"41194443","id":"PMC_41194443","title":"CRISPR/Cas9-mediated genome editing reveals six testis-enriched genes dispensable for male fertility in mice.","date":"2025","source":"Journal of biomedical research","url":"https://pubmed.ncbi.nlm.nih.gov/41194443","citation_count":0,"is_preprint":false},{"pmid":"41614567","id":"PMC_41614567","title":"The 129S1/SvImJ Mouse Strain Is Resistant to the Effects of Early Embryonic Alcohol and Other Sonic Hedgehog Inhibitors.","date":"2026","source":"Birth defects research","url":"https://pubmed.ncbi.nlm.nih.gov/41614567","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":8073,"output_tokens":1788,"usd":0.02552,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":8625,"output_tokens":2488,"usd":0.052663,"stage2_stop_reason":"end_turn"},"total_usd":0.078183,"stage1_batch_id":"msgbatch_01AsR7g8dGTEiEw9U83bJeWn","stage2_batch_id":"msgbatch_01YV5ZyxtFNUZEjxRcpd9jzb","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2014,\n      \"finding\": \"EFCAB7 forms a complex with IQCE that anchors the EVC-EVC2 heterodimer in a signaling microdomain at the base of primary cilia (the EvC zone). EFCAB7 directly binds a C-terminal disordered region of EVC2 that is deleted in Weyers syndrome patients. Depletion of EFCAB7 causes mislocalization of EVC-EVC2 within cilia and impairs activation of the transcription factor GLI2, phenocopying the Weyers cellular phenotype.\",\n      \"method\": \"Co-immunoprecipitation, ciliary localization by immunofluorescence, siRNA knockdown with GLI2 activation assay, evolutionary/domain analysis\",\n      \"journal\": \"Developmental Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, direct localization experiments, loss-of-function with defined pathway phenotype (GLI2 activation), binding domain mapping; foundational paper with 77 citations and multiple orthogonal methods\",\n      \"pmids\": [\"24582806\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"The EvC zone targeting of EVC-EVC2 depends on two separate EFCAB7-binding motifs within EVC2's Weyers-deleted peptide; only one had been previously characterized. SUMOylation of EVC-EVC2 cytosolic tails with SUMO3 enhances complex accumulation at the EvC zone, possibly via increased binding to the EFCAB7-IQCE complex. EFCAB7 and IQCE were confirmed as EVC interactors by endogenous EVC protein interactome profiling in control and Evc-null cells.\",\n      \"method\": \"Endogenous Co-IP/mass spectrometry (interactome screen in Evc-null vs. control cells), SUMOylation assays, ciliary localization by immunofluorescence, mapping of EVC2 binding motifs\",\n      \"journal\": \"Frontiers in Cell and Developmental Biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — endogenous interactome confirmed by MS and Co-IP, SUMOylation functional assay, motif mapping; single lab, multiple orthogonal methods\",\n      \"pmids\": [\"37576597\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"A splicing variant (c.683-1G>C) in EFCAB7 causes exon skipping and production of a non-functional protein. Mice carrying this variant exhibit impaired ciliogenesis, disrupted Hedgehog/Shh-Gli pathway activity, and transcriptional downregulation of Gli target genes (Myh6, Zfpm1, Nkx2-5), resulting in abnormal cardiac development including Tetralogy of Fallot-like defects.\",\n      \"method\": \"Sanger sequencing of patients, in vitro and in vivo splicing assays, mouse model with variant knock-in, ciliogenesis assay, CUT&Tag for Gli binding, bulk RNA-seq of embryonic hearts\",\n      \"journal\": \"The Journal of Biological Chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo mouse model with multiple readouts (ciliogenesis, pathway activity, transcriptomics), single lab, multiple orthogonal methods\",\n      \"pmids\": [\"39894222\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Full deletion of Efcab7 in mice sensitizes fetuses to craniofacial and ocular malformations induced by the Smo antagonist vismodegib or by alcohol, while reducing malformations induced by the Smo agonist SAG. This establishes EFCAB7 as a positive regulator of Smoothened (Smo) activity within the Shh pathway in vivo.\",\n      \"method\": \"CRISPR/genetic knockout mouse model, pharmacological challenge with Smo agonist/antagonist and alcohol at gastrulation stage, fetal phenotypic scoring\",\n      \"journal\": \"Reproductive Toxicology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KO mouse with defined epistatic relationship to Smo agonist/antagonist, single lab, genetic and pharmacological dissection\",\n      \"pmids\": [\"39366525\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"EFCAB7 directly interacts with PARK7 (DJ-1) in hepatocellular carcinoma cells, as demonstrated by mass spectrometry and Co-IP. Overexpression of PARK7 in EFCAB7-knockdown cells rescues tumor cell proliferation and metastasis, indicating EFCAB7 acts upstream of PARK7 in a cancer cell context.\",\n      \"method\": \"Mass spectrometry, Co-immunoprecipitation, siRNA knockdown, PARK7 rescue overexpression, in vitro proliferation/metastasis assays\",\n      \"journal\": \"Aging\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single Co-IP and MS from one lab, no structural or reconstitution validation, context is cancer cell line only\",\n      \"pmids\": [\"39276379\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"CRISPR/Cas9-mediated homozygous deletion of Efcab7 in mice produces no detectable defects in male fertility, sperm motility, or testis/epididymis morphology, indicating Efcab7 is dispensable for male reproduction when ablated alone.\",\n      \"method\": \"CRISPR/Cas9 knockout mice, mating tests, sperm motility analysis, histological phenotyping\",\n      \"journal\": \"Journal of Biomedical Research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — clean KO with multiple phenotypic readouts confirming null result; single lab, single study, but rigorous in vivo approach\",\n      \"pmids\": [\"41194443\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"EFCAB7 is a ciliary protein that forms a complex with IQCE to anchor the EVC-EVC2 heterodimer at the EvC zone (base of primary cilia), thereby positively regulating Hedgehog/Smoothened-GLI2 signaling; its loss mislocalizes EVC-EVC2, impairs GLI2 activation, and causes developmental defects including skeletal anomalies, polydactyly, and congenital heart defects, while SUMOylation of EVC-EVC2 and two distinct EFCAB7-binding motifs on EVC2 further modulate complex stability and ciliary targeting.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"EFCAB7 is a primary cilium protein that functions as a positive regulator of Hedgehog signaling by anchoring the EVC-EVC2 heterodimer within a specialized signaling microdomain at the ciliary base (the EvC zone) [#0]. Together with IQCE, EFCAB7 forms a complex that directly binds a C-terminal disordered region of EVC2 — the same region deleted in Weyers syndrome — and its depletion mislocalizes EVC-EVC2 within cilia and impairs activation of the GLI2 transcription factor [#0]. EvC zone targeting depends on two distinct EFCAB7-binding motifs in the EVC2 Weyers-deleted peptide, and SUMOylation of the EVC-EVC2 cytosolic tails enhances complex accumulation at this site, consistent with modulation of binding to the EFCAB7-IQCE complex [#1]. In vivo, EFCAB7 acts upstream as a positive regulator of Smoothened activity: full deletion in mice sensitizes fetuses to malformations induced by Smo antagonism and protects against Smo agonism [#3], and a disease-associated splicing variant disrupts ciliogenesis, downregulates Gli target genes (Myh6, Zfpm1, Nkx2-5), and produces Tetralogy of Fallot-like cardiac defects [#2]. EFCAB7 loss is nonetheless dispensable for male fertility and sperm motility when ablated alone [#5].\",\n  \"teleology\": [\n    {\n      \"year\": 2014,\n      \"claim\": \"Established how EVC-EVC2 is retained at the ciliary base and linked EFCAB7 to a defined Hedgehog signaling output, answering how a Weyers-syndrome–associated EVC2 region is functionally read out.\",\n      \"evidence\": \"Co-IP, ciliary immunofluorescence, EFCAB7 siRNA knockdown with GLI2 activation assay, and binding-domain mapping in mammalian cells\",\n      \"pmids\": [\"24582806\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not resolve the structural basis of the EFCAB7-IQCE-EVC2 complex\", \"Stoichiometry and assembly order at the EvC zone not defined\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Refined the molecular interface and regulation, showing EvC zone targeting requires two separate EFCAB7-binding motifs and is enhanced by SUMOylation of the EVC-EVC2 tails.\",\n      \"evidence\": \"Endogenous EVC interactome by Co-IP/MS in Evc-null vs control cells, SUMOylation assays, ciliary localization, and EVC2 motif mapping (single lab)\",\n      \"pmids\": [\"37576597\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which SUMO3 increases EFCAB7-IQCE binding not directly demonstrated\", \"SUMO ligase/site responsible not identified\", \"Single lab\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Placed EFCAB7 epistatically within the Shh pathway in vivo as a positive regulator of Smoothened activity, distinguishing its role from upstream/downstream nodes.\",\n      \"evidence\": \"CRISPR Efcab7-knockout mice challenged with Smo agonist (SAG), Smo antagonist (vismodegib), or alcohol at gastrulation, with fetal phenotypic scoring\",\n      \"pmids\": [\"39366525\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Does not define the molecular step at which EFCAB7 modulates Smo\", \"Single lab\", \"Epistasis inferred pharmacologically rather than biochemically\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Connected an EFCAB7 loss-of-function variant to a human-relevant developmental disease mechanism, tying ciliogenesis and Gli-target transcription to cardiac morphogenesis.\",\n      \"evidence\": \"Splicing assays, variant knock-in mouse, ciliogenesis assay, CUT&Tag for Gli binding, and bulk RNA-seq of embryonic hearts\",\n      \"pmids\": [\"39894222\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct effect of EFCAB7 on ciliogenesis machinery not mechanistically resolved\", \"Single lab\", \"Causality between Gli-target downregulation and specific cardiac defects not fully dissected\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Reported a non-ciliary context in which EFCAB7 interacts with PARK7 and acts upstream of it to support tumor cell proliferation and metastasis.\",\n      \"evidence\": \"Mass spectrometry and Co-IP in hepatocellular carcinoma cells with siRNA knockdown and PARK7 rescue overexpression (in vitro)\",\n      \"pmids\": [\"39276379\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single Co-IP and MS from one lab, no reciprocal or structural validation\", \"Restricted to one cancer cell line context\", \"Relationship to ciliary function unknown\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Defined the boundary of EFCAB7 requirement in vivo, showing it is dispensable for male reproduction when ablated alone.\",\n      \"evidence\": \"CRISPR/Cas9 knockout mice with mating tests, sperm motility analysis, and histological phenotyping\",\n      \"pmids\": [\"41194443\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Possible redundancy with paralogs not tested\", \"Female reproductive phenotype not addressed\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The structural architecture of the EFCAB7-IQCE-EVC2 assembly and the precise molecular step at which EFCAB7 enables Smoothened-to-GLI2 signal transduction remain undefined.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structure of the EFCAB7-IQCE-EVC2 complex\", \"Direct biochemical link between complex anchoring and Smo activation unresolved\", \"PARK7 interaction not integrated with ciliary role\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 2, 3]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [2, 3]}\n    ],\n    \"complexes\": [\"EFCAB7-IQCE complex\", \"EvC zone (EVC-EVC2-EFCAB7-IQCE)\"],\n    \"partners\": [\"IQCE\", \"EVC2\", \"EVC\", \"PARK7\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}