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Fine-tuning of the GRN components underlies facial shape variation, yet how those networks in the midface are connected and activated remain poorly understood. Here, we show that concerted inactivation of Tfap2a and Tfap2b in the murine neural crest, even during the late migratory phase, results in a midfacial cleft and skeletal abnormalities. Bulk and single-cell RNA-seq profiling reveal that loss of both TFAP2 family members dysregulates numerous midface GRN components involved in midface morphogenesis, patterning and differentiation. Notably, Alx1, Alx3 and Alx4 (ALX) transcript levels are reduced, whereas ChIP-seq analyses suggest TFAP2 family members directly and positively regulate ALX gene expression. Tfap2a, Tfap2b and ALX co-expression in midfacial neural crest cells of both mouse and zebrafish implies conservation of this regulatory axis across vertebrates. Consistent with this notion, tfap2a zebrafish mutants present with abnormal alx3 expression patterns, Tfap2a binds ALX loci and tfap2a-alx3 genetic interactions are observed. Together, these data demonstrate TFAP2 paralogs regulate vertebrate midfacial development in part by activating expression of ALX transcription factor genes.<\/jats:p>","DOI":"10.1242\/dev.202095","type":"journal-article","created":{"date-parts":[[2023,12,8]],"date-time":"2023-12-08T15:44:05Z","timestamp":1702050245000},"source":"Crossref","is-referenced-by-count":16,"title":["TFAP2 paralogs regulate midfacial development in part through a conserved ALX genetic pathway"],"prefix":"10.1242","volume":"151","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0979-1286","authenticated-orcid":false,"given":"Timothy T.","family":"Nguyen","sequence":"first","affiliation":[{"name":"Iowa Institute for Oral Health Research, College of Dentistry and Dental Clinics, University of Iowa 1 , Iowa City, IA 52242 , USA"},{"name":"College of Dentistry and Dental Clinics, University of Iowa 2 Department of Periodontics , , Iowa City, IA 52242 , USA"},{"name":"Carver College of Medicine, University of Iowa 3 Department of Anatomy and Cell Biology , , Iowa City, IA 52242 , USA"},{"name":"University of Iowa 4 Interdisciplinary Graduate Program in Genetics , , Iowa City, IA 52242 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4222-5235","authenticated-orcid":false,"given":"Jennyfer M.","family":"Mitchell","sequence":"additional","affiliation":[{"name":"University of Colorado Anschutz Medical Campus 5 Department of Craniofacial Biology , , Aurora, CO 80045 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michaela D.","family":"Kiel","sequence":"additional","affiliation":[{"name":"Iowa Institute for Oral Health Research, College of Dentistry and Dental Clinics, University of Iowa 1 , Iowa City, IA 52242 , USA"},{"name":"College of Dentistry and Dental Clinics, University of Iowa 2 Department of Periodontics , , Iowa City, IA 52242 , USA"},{"name":"Carver College of Medicine, University of Iowa 3 Department of Anatomy and Cell Biology , , Iowa City, IA 52242 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8926-4413","authenticated-orcid":false,"given":"Colin P.","family":"Kenny","sequence":"additional","affiliation":[{"name":"Carver College of Medicine 6 Department of Surgery , , , Iowa City, IA 52242 , USA"},{"name":"University of Iowa 6 Department of Surgery , , , Iowa City, IA 52242 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hong","family":"Li","sequence":"additional","affiliation":[{"name":"University of Colorado Anschutz Medical Campus 5 Department of Craniofacial Biology , , Aurora, CO 80045 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kenneth L.","family":"Jones","sequence":"additional","affiliation":[{"name":"University of Colorado Anschutz Medical Campus, Children's Hospital Colorado 7 Department of Pediatrics , , Aurora, CO 80045 , 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