Suchergebnisse

Regulation of male sexual differentiation by a Y chromosome-linked male determining factor (M-factor) is one of a diverse array of sex determination mechanisms found in insects. By deep sequencing of small RNAs from Bactrocera dorsalis early embryos, we identified an autosomal-derived microRNA, miR-1-3p, that has predicted target sites in the transformer gene (Bdtra) required for female sex determination. We further demonstrate by both in vitro and in vivo tests that miR-1-3p suppresses Bdtra expression. Injection of a miR-1-3p mimic in early embryos results in 87-92% phenotypic males, whereas knockdown of miR-1-3p by an inhibitor results in 67-77% phenotypic females. Finally, CRISPR/Cas9-mediated knockout of miR-1-3p results in the expression of female-specific splice variants of Bdtra and doublesex (Bddsx), and induced sex reversal of XY individuals into phenotypic females. These results indicate that miR-1-3p is required for male sex determination in early embryogenesis in B. dorsalis as an intermediate male determiner. ; National Key R&D Program of China [2017YFD0202000]; earmarked fund for the China Agricultural Research System [CARS-26]; International Atomic Energy Agency's Coordinated Research Project [D42016]; National Natural Science Foundation of ChinaNational Natural Science Foundation of China (NSFC) [31872931, 31572008]; Fundamental Research Funds for the Central UniversitiesFundamental Research Funds for the Central Universities [2662019PY055]; USDA-NIFA-AFRI Foundational Program [2016-67013-25087]; FFABR2017; NIHUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USA [AI123338, AI121284] ; This research was supported by National Key R&D Program of China (No. 2017YFD0202000), the earmarked fund for the China Agricultural Research System (No. CARS-26), International Atomic Energy Agency's Coordinated Research Project (No. D42016), the National Natural Science Foundation of China (Nos. 31872931 and 31572008) and the Fundamental Research Funds for the Central Universities (No. 2662019PY055) to H.Z. Support is also acknowledged from the USDA-NIFA-AFRI Foundational Program (No. 2016-67013-25087) to AMH, grant FFABR2017 to G.S., and NIH grants AI123338 and AI121284 to Z.J.T. We thank Dr. Eric Jang from USDA-ARS, Pacific West Area, Tropical Crop and Commodity Protection Research Unit, for providing the B. dorsalis white/brown pupae sexing strain. ; Public domain authored by a U.S. government employee