{"doi":"10.1172/jci.insight.171661","title":"Generating endogenous Myh11-driven Cre mice for sex-independent gene deletion in smooth muscle cells","abstract":"Specific and efficient smooth muscle cell-targeted (SMC-targeted) gene deletion is typically achieved by pairing SMMHC-CreERT2-Tg mice with mice carrying the loxP-flanked gene. However, the transgene, CreERT2, is not controlled by the endogenous Myh11 gene promoter, and the codon-modified iCreERT2 exhibits significant tamoxifen-independent leakage. Furthermore, because the Cre-bearing bacterial artificial chromosome (BAC) is inserted onto the Y chromosome, the SMMHC-CreERT2-Tg mice strain can only exhibit gene deletions in male mice. Additionally, there is a lack of Myh11-driven constitutive Cre mice when tamoxifen usage is a concern. We used CRISPR/Cas9-mediated homologous recombination between a donor vector carrying the CreNLSP2A or CreERT2-P2A sequence and homologous arm surrounding the translation start site of the Myh11 gene to generate Cre-knockin mice. The P2A sequence enables the simultaneous translation of Cre and endogenous proteins. Using reporter mice, we assessed Cre-mediated recombination efficiency, specificity, tamoxifen-dependent controllability, and functionality in both sexes. Both constitutive (Myh11-CreNLSP2A) and inducible (Myh11-CreERT2-P2A) Cre mice demonstrated efficient, SMC-specific, sex-independent Cre recombinase activity without confounding endogenous gene expression. Combined with recently generated BAC transgenic Myh11-CreERT2-RAD mice and the Itga8-CreERT2 mouse models, our models will help expand the research toolbox, facilitating unbiased and comprehensive research in SMCs and SMC-dependent cardiovascular diseases.","journal":"JCI Insight","year":2023,"id":360762,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":9,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9491,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":243090,"name":"Guizhen Zhao","orcid":"0000-0002-2648-8986","position":1,"is_corresponding":false},{"id":244203,"name":"Ziyi Chang","orcid":null,"position":2,"is_corresponding":false},{"id":243093,"name":"Tianqing Zhu","orcid":"0000-0003-3592-5276","position":3,"is_corresponding":false},{"id":1112569,"name":"Ying Zhao","orcid":"0000-0002-5511-6284","position":4,"is_corresponding":false},{"id":243091,"name":"Haocheng Lu","orcid":"0000-0002-5740-8010","position":5,"is_corresponding":false},{"id":858168,"name":"Chao Xue","orcid":"0000-0002-6528-3975","position":6,"is_corresponding":false},{"id":317594,"name":"Thomas L. Saunders","orcid":"0000-0003-2015-101X","position":7,"is_corresponding":false},{"id":243095,"name":"Yanhong Guo","orcid":"0000-0002-3334-9159","position":8,"is_corresponding":false},{"id":243094,"name":"Lin Chang","orcid":"0000-0002-5621-8426","position":9,"is_corresponding":false},{"id":237553,"name":"Y. Eugene Chen","orcid":"0000-0003-2357-7825","position":10,"is_corresponding":false},{"id":243097,"name":"Jifeng Zhang","orcid":"0000-0001-5161-4705","position":11,"is_corresponding":false},{"id":243092,"name":"Yang Zhao","orcid":"0000-0002-1428-5145","position":0,"is_corresponding":true}],"reference_count":55,"raw_metadata":null,"created_at":"2026-07-19T01:14:06.433417Z","pmid":"37289544","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}