title: Direct production of XY(DMY-) sex reversal female medaka (Oryzias latipes) by embryo microinjection of TALENs.
abstract: Medaka is an ideal model for sex determination and sex reversal, such as XY phenotypically female patients in humans. Here, we assembled improved TALENs targeting the DMY gene and generated XY(DMY-) mutants to investigate gonadal dysgenesis in medaka. DMY-TALENs resulted in indel mutations at the targeted loci (46.8%). DMY-nanos3UTR-TALENs induced mutations were passed through the germline to F1 generation with efficiencies of up to 91.7%. XY(DMY-) mutants developed into females, laid eggs, and stably passed the Y(DMY-) chromosome to next generation. RNA-seq generated 157 million raw reads from WT male (WT_M_TE), WT female (WT_F_OV) and XY(DMY-) female medaka (TA_F_OV) gonad libraries. Differential expression analysis identified 144 up- and 293 down-regulated genes in TA_F_OV compared with WT_F_OV, 387 up- and 338 down-regulated genes in TA_F_OV compared with WT_M_TE. According to genes annotation and functional prediction, such as Wnt1 and PRCK, it revealed that incomplete ovarian function and reduced fertility of XY(DMY-) mutant is closely related to the wnt signaling pathway. Our results provided the transcriptional profiles of XY(DMY-) mutants, revealed the mechanism between sex reversal and DMY in medaka, and suggested that XY(DMY-) medaka was a novel mutant that is useful for investigating gonadal dysgenesis in phenotypic female patients with the 46, XY karyotype.
authors.affiliation: Department of Genetics, School of Basic Medical Sciences, Wuhan University, Wuhan, Hubei, P. R. China, 430071.|State Key Laboratory of Freshwater Ecology and Biotechnology, Institutes of Hydrobiology, Chinese Academy of Sciences, Wuhan, Hubei, P. R. China, 430072.
authors.forename: Daji
authors.initials: D
authors.lastname: Luo
authors.affiliation: School of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong, P. R. China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Freshwater Ecology and Biotechnology, Institutes of Hydrobiology, Chinese Academy of Sciences, Wuhan, Hubei, P. R. China, 430072.
authors.forename: Ji
authors.initials: J
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Freshwater Ecology and Biotechnology, Institutes of Hydrobiology, Chinese Academy of Sciences, Wuhan, Hubei, P. R. China, 430072.
authors.forename: Xiaoqin
authors.initials: X
authors.lastname: Xia
authors.affiliation: State Key Laboratory of Freshwater Ecology and Biotechnology, Institutes of Hydrobiology, Chinese Academy of Sciences, Wuhan, Hubei, P. R. China, 430072.
authors.forename: Mengxi
authors.initials: M
authors.lastname: Cao
authors.affiliation: Department of Genetics, School of Basic Medical Sciences, Wuhan University, Wuhan, Hubei, P. R. China, 430071.
authors.forename: Bin
authors.initials: B
authors.lastname: Cheng
authors.affiliation: Department of Genetics, School of Basic Medical Sciences, Wuhan University, Wuhan, Hubei, P. R. China, 430071.
authors.forename: Xuejuan
authors.initials: X
authors.lastname: Wang
authors.affiliation: Lillehei Heart Institute, University of Minnesota, Minneapolis, USA.
authors.forename: Wuming
authors.initials: W
authors.lastname: Gong
authors.affiliation: Department of Genetics, School of Basic Medical Sciences, Wuhan University, Wuhan, Hubei, P. R. China, 430071.
authors.forename: Chao
authors.initials: C
authors.lastname: Qiu
authors.affiliation: State Key Laboratory of Freshwater Ecology and Biotechnology, Institutes of Hydrobiology, Chinese Academy of Sciences, Wuhan, Hubei, P. R. China, 430072.
authors.forename: Yunsheng
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: School of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong, P. R. China.
authors.forename: Christopher Hon Ki
authors.initials: CH
authors.lastname: Cheng
authors.affiliation: State Key Laboratory of Freshwater Ecology and Biotechnology, Institutes of Hydrobiology, Chinese Academy of Sciences, Wuhan, Hubei, P. R. China, 430072.
authors.forename: Zuoyan
authors.initials: Z
authors.lastname: Zhu
authors.affiliation: State Key Laboratory of Freshwater Ecology and Biotechnology, Institutes of Hydrobiology, Chinese Academy of Sciences, Wuhan, Hubei, P. R. China, 430072.
authors.forename: Wei
authors.initials: W
authors.lastname: Hu
mesh_terms: D000818:Animals; D001483:Base Sequence; D003851:Deoxyribonucleases / Q000378:metabolism*; D004625:Embryo, Nonmammalian / Q000378:metabolism; D005260:Female; D005854:Germ Cells / Q000166:cytology / Q000378:metabolism; D006066:Gonads / Q000378:metabolism; D059014:High-Throughput Nucleotide Sequencing; D008297:Male; D008845:Microinjections; D009154:Mutation; D009990:Oryzias / Q000235:genetics* / Q000254:growth & development; D010641:Phenotype; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification; D017423:Sequence Analysis, RNA; D012730:Sex Chromosomes; D019849:Sex Determination Processes
chemical_list: D012313:RNA; D003851:Deoxyribonucleases