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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.

Direct production of XYDMY− sex reversal female medaka (Oryzias latipes) by embryo microinjection of TALENs