{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,22]],"date-time":"2026-04-22T06:53:23Z","timestamp":1776840803714,"version":"3.51.2"},"reference-count":31,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,4,22]],"date-time":"2026-04-22T00:00:00Z","timestamp":1776816000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell Dev. Biol."],"abstract":"\n Background<\/jats:title>\n \n We previously demonstrated that fibroblasts can be reprogrammed through a proliferative progenitor stage to drive both cardiomyogenesis and neovascularization. However, it remains to be determined whether cardiac fibroblasts (CFs), the primary mediators of post-injury remodeling, retain the plasticity to be concurrently redirected into cardiovascular lineages within the\n in vivo<\/jats:italic>\n environment.\n <\/jats:p>\n <\/jats:sec>\n \n Methods<\/jats:title>\n \n This study aimed to activate endogenous cardiogenic genes (\n Gata4<\/jats:italic>\n ,\n Nkx2<\/jats:italic>\n .5,\n Tbx5<\/jats:italic>\n ,\n Isl1<\/jats:italic>\n , and\n Smarcd3<\/jats:italic>\n , referred to as\n GNTIS<\/jats:italic>\n ) in CFs using a CRISPR activation (CRISPRa) system. CFs isolated from fibroblast-specific transgenic dCas9 mice were used to validate cardiogenic gene activation mediated by single-guide RNAs and VP64 activators delivered\n via<\/jats:italic>\n adeno-associated viral vectors (AAVs). The reprogramming potential of CFs into cardiovascular lineages was further evaluated in dCas9 mice subjected to myocardial infarction followed by administration of pooled AAVs.\n <\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n Individual\n GNTIS<\/jats:italic>\n sgRNAs effectively upregulated their respective targets in the transfected CFs. The pooled sgRNA\n \n GNTIS<\/jats:italic>\n <\/jats:sup>\n induced cardiac-like phenotypes in the CFs, as demonstrated by the Nkx2.5 cardiac progenitor reporter and increased cardiac differentiation markers. Subsequently, AAV-sgRNA\n \n GNTIS<\/jats:italic>\n <\/jats:sup>\n was assessed\n in vivo<\/jats:italic>\n in dCas9 mice following myocardial infarction. While global cardiac function did not reach statistical significance,\n GNTIS<\/jats:italic>\n activation improved key hemodynamic parameters, effectively preserving cardiac performance compared to controls. Immunostaining further revealed that\n GNTIS<\/jats:italic>\n -activated CFs exhibited the potential for transdifferentiation into cardiomyocyte-like or vascular smooth muscle cell-like lineages within the infarcted heart, which was not observed in the control group.\n <\/jats:p>\n <\/jats:sec>\n \n Conclusion<\/jats:title>\n \n This CRISPRa transgenic model provides a foundational proof of concept for\n in vivo<\/jats:italic>\n reprogramming of CFs into cardiovascular cells.\n <\/jats:p>\n <\/jats:sec>","DOI":"10.3389\/fcell.2026.1812941","type":"journal-article","created":{"date-parts":[[2026,4,22]],"date-time":"2026-04-22T05:54:41Z","timestamp":1776837281000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["In vivo CRISPR-mediated activation of cardiogenic genes to reprogram cardiac fibroblasts"],"prefix":"10.3389","volume":"14","author":[{"given":"Suchandrima","family":"Dutta","sequence":"first","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, College of Medicine, University of Cincinnati","place":["Cincinnati, OH, United States"]},{"name":"Department of Internal Medicine, College of Medicine, University of Cincinnati","place":["Cincinnati, OH, United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sophie","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, College of Medicine, University of Cincinnati","place":["Cincinnati, OH, United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Waqas","family":"Ahmad","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, College of Medicine, University of Cincinnati","place":["Cincinnati, OH, United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wei","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, College of Medicine, University of Cincinnati","place":["Cincinnati, OH, United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yigang","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, College of Medicine, University of Cincinnati","place":["Cincinnati, OH, United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jialiang","family":"Liang","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, College of Medicine, University of Cincinnati","place":["Cincinnati, OH, United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2026,4,22]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"940","DOI":"10.3390\/bioengineering12090940","article-title":"In vivo targeted reprogramming of cardiac fibroblasts for heart regeneration: advances and therapeutic potential","volume":"12","author":"Ahmad","year":"2025","journal-title":"Bioeng. 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