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Aromatase encoded by Cyp19a1 is the key enzyme that catalyzes the conversion of androgen to estrogen. Correlative analyses implicate the potential involvement of aromatase in reptilian sexual development, however, the direct genetic evidence is lacking. Herein, we found that Cyp19a1 exhibited temperature-dependent sexually dimorphic expression, and located in the medullary somatic cells in early female embryos of the red-eared slider turtle (Trachemys scripta elegans), before the gonad is distinct. To determine the functional role of Cyp19a1 in turtle ovarian determination, we established loss- and gain-of-function models through in ovo lentivirus-mediated genetic manipulation. At female-producing temperature, inhibition of aromatase or knockdown of Cyp19a1 in turtle embryos resulted in female-to-male sex reversal, with the formation of a testis-like structure and a male distribution pattern of germ cells, as well as ectopic expression of male-specific markers (SOX9 and AMH) and disappearance of ovarian regulator FOXL2. On the contrary, overexpression of Cyp19a1 at male-producing temperature led to male-to-female sex reversal. In conclusion, our results suggest that Cyp19a1 is both necessary and sufficient for ovarian determination in the red-eared slider turtle, establishing causality and a direct genetic link between aromatase and reptilian sex determination and differentiation.<\/jats:p>","DOI":"10.1093\/genetics\/iyad041","type":"journal-article","created":{"date-parts":[[2023,3,22]],"date-time":"2023-03-22T19:15:10Z","timestamp":1679512510000},"source":"Crossref","is-referenced-by-count":9,"title":["Loss- and gain-of-function analyses reveal the essential role of <i>Cyp19a1<\/i> in ovarian determination of the red-eared slider turtle"],"prefix":"10.1093","volume":"224","author":[{"given":"Jiadong","family":"Shen","sequence":"first","affiliation":[{"name":"Institute of Animal Sex and Development, Zhejiang Wanli University , Ningbo 315100 , China"},{"name":"College of Biological and Environmental Sciences, Zhejiang Wanli University , Ningbo 315100 , 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Ningbo 315100 , China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiandong","family":"Lei","sequence":"additional","affiliation":[{"name":"Institute of Animal Sex and Development, Zhejiang Wanli University , Ningbo 315100 , China"},{"name":"College of Biological and Environmental Sciences, Zhejiang Wanli University , Ningbo 315100 , China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qinglu","family":"Ma","sequence":"additional","affiliation":[{"name":"Institute of Animal Sex and Development, Zhejiang Wanli University , Ningbo 315100 , China"},{"name":"College of Biological and Environmental Sciences, Zhejiang Wanli University , Ningbo 315100 , China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ling","family":"Xiao","sequence":"additional","affiliation":[{"name":"Institute of Animal Sex and Development, Zhejiang Wanli University , Ningbo 315100 , China"},{"name":"College of Biological and Environmental Sciences, Zhejiang Wanli University , Ningbo 315100 , China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7978-6629","authenticated-orcid":false,"given":"Chutian","family":"Ge","sequence":"additional","affiliation":[{"name":"Institute of Animal Sex and Development, Zhejiang Wanli University , Ningbo 315100 , China"},{"name":"College of Biological and Environmental Sciences, Zhejiang Wanli University , Ningbo 315100 , China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2023,3,22]]},"reference":[{"issue":"5","key":"2024091718035431900_iyad041-B1","doi-asserted-by":"publisher","first-page":"490","DOI":"10.1002\/jez.1092","article-title":"Sex reversal and aromatase in the European pond turtle: treatment with letrozole after the thermosensitive period for sex determination","volume":"290","author":"Belaid","year":"2001","journal-title":"J Exp 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