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

Daji Luo, Yun Liu, Ji Chen, Xiaoqin Xia, Mengxi Cao, Bin Cheng, Xuejuan Wang, Wuming Gong, Chao Qiu, Yunsheng Zhang, Christopher Hon Ki Cheng, Zuoyan Zhu, Wei Hu

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

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%. XYDMY- mutants developed into females, laid eggs, and stably passed the YDMY-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 XYDMY- 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.

Original languageEnglish (US)
Article number14057
JournalScientific reports
Volume5
DOIs
StatePublished - Sep 14 2015

Bibliographical note

Funding Information:
We thank Ms. Li Ming (Institute of Hydrobiology, Chinese Academy of Sciences) for her kind help during the feeding of medaka and microinjection of medaka embryos. This work was supported by the National Science Foundation for Distinguished Young Scholars of China (31325026 to WH), the National High Technology Research and Development Program of China (2011AA100404 to WH), the National Science Foundation of China (31472263 to DL), and the Chinese Academy of Sciences (XDA08010106 and 2011FBZ19 to WH).

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