title: RNAi as a tool to control the sex ratio of mouse offspring by interrupting Zfx/Zfy genes in the testis.
abstract: The objective of this study was to explore a novel method to alter the sex-ratio balance of mouse offspring by silencing the paralogous genes Zfx/Zfy (Zinc finger X/Y-chromosomal transcription factor gene) during spermatogenesis. Four recombined vectors PRZ1, PRZ2, PRZ3, and PRZ4 (RNAi-Ready-pSIREN-RetroQ-ZsGreen) were constructed for interrupting the Zfx gene. Additionally, a recombined vector Psilencer/Zfy-shRNA was constructed for interrupting the Zfy gene. Male mice were randomly divided into 8 groups, with 20 animals per group. Five groups of mice were injected with PRZ1, PRZ2, PRZ3, PRZ4, and Psilencer/Zfy-shRNA vectors, respectively. The three control groups were injected with an equal volume of physiological saline, empty RNAi-Ready-pSIREN-RetroQ-ZsGreen vector, and empty Psilencer/Zfy-shRNA vector, respectively. All groups were injected every 7 days for a total of four injections. Fourteen days after the fourth injection, 10 male mice from each group were mated individually with 10 females. Testicular tissue of 10 male mice in each group was collected, and the expression level of Zfx/Zfy mRNA was determined by qRT-PCR. Results showed that, compared with the empty RNAi-Ready-pSIREN-RetroQ-ZsGreen vector and the physiological saline group, expression of Zfx mRNA decreased significantly after injection of PRZ1 (p < 0.01), PRZ3 (p < 0.01), and PRZ4 (p < 0.01), and 78.75 ± 7.50% of the offspring were male in PRZ4 group, significantly higher than the offspring derived from the empty RNAi-Ready-pSIREN-RetroQ-ZsGreen vector and physiological saline group (p < 0.01). In the PRZ1 group, the expression of Zfx mRNA was also significantly lower (p < 0.01), but the male rate of offspring was not different (p > 0.05). Conversely, the expression of Zfy mRNA decreased significantly after injection of Psilencer/Zfy-shRNA (p < 0.01) and 31.00 ± 11.00% of the offspring were male, significantly lower than in the physiological saline group (p < 0.01). In conclusion, our findings show that RNAi-mediated disruption of Zfx/Zfy in mouse testis affected X/Y spermatogenesis. Additionally, results suggest that the paralogous genes Zfx/Zfy play an important role in the process of X and Y sperm development. The individual interference of Zfx/Zfy may predict the outcome of X and Y haploid sperms. Presented herein is an advanced method developed to control mouse X/Y spermatogenesis and sex ratio of offspring.
authors.affiliation: College of Animal Science and Technology, Shihezi University, Xinjiang, Xinjiang Uygur Autonomous Region, China.
authors.forename: YongSheng
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: College of Animal Science and Technology, Shihezi University, Xinjiang, Xinjiang Uygur Autonomous Region, China.|Xinjiang Agricultural Vocational Technical College, Shihezi, China.
authors.forename: JiFeng
authors.initials: J
authors.lastname: Xi
authors.affiliation: College of Animal Science and Technology, Shihezi University, Xinjiang, Xinjiang Uygur Autonomous Region, China. jiabin@shzu.edu.cn.
authors.forename: Bin
authors.initials: B
authors.lastname: Jia
authors.affiliation: College of Animal Science and Technology, Shihezi University, Xinjiang, Xinjiang Uygur Autonomous Region, China.|Xinjiang Agricultural Vocational Technical College, Shihezi, China.
authors.forename: XiangZu
authors.initials: X
authors.lastname: Wang
authors.affiliation: College of Animal Science and Technology, Shihezi University, Xinjiang, Xinjiang Uygur Autonomous Region, China.
authors.forename: XuHai
authors.initials: X
authors.lastname: Wang
authors.affiliation: College of Animal Science and Technology, Shihezi University, Xinjiang, Xinjiang Uygur Autonomous Region, China.
authors.forename: ChaoCheng
authors.initials: C
authors.lastname: Li
authors.affiliation: College of Animal Science and Technology, Shihezi University, Xinjiang, Xinjiang Uygur Autonomous Region, China.
authors.forename: YaQiang
authors.initials: Y
authors.lastname: Li
authors.affiliation: College of Animal Science and Technology, Shihezi University, Xinjiang, Xinjiang Uygur Autonomous Region, China.
authors.forename: XianCun
authors.initials: X
authors.lastname: Zeng
authors.affiliation: College of Animal Science and Technology, Shihezi University, Xinjiang, Xinjiang Uygur Autonomous Region, China.
authors.forename: RuiWen
authors.initials: R
authors.lastname: Ying
authors.affiliation: College of Life Sciences, Shihezi University, Xinjiang, Xinjiang Uygur Autonomous Region, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.affiliation: College of Animal Science and Technology, Shihezi University, Xinjiang, Xinjiang Uygur Autonomous Region, China.
authors.forename: Song
authors.initials: S
authors.lastname: Jiang
authors.affiliation: College of Animal Science and Technology, Shihezi University, Xinjiang, Xinjiang Uygur Autonomous Region, China.
authors.forename: FangYuan
authors.initials: F
authors.lastname: Yuan
mesh_terms: D000818:Animals; D004268:DNA-Binding Proteins / Q000235:genetics*; D005260:Female; D020868:Gene Silencing; D051741:Kruppel-Like Transcription Factors / Q000235:genetics*; D008297:Male; D051379:Mice; D034622:RNA Interference; D012744:Sex Ratio; D013091:Spermatogenesis / Q000235:genetics*; D013094:Spermatozoa / Q000254:growth & development / Q000378:metabolism; D013737:Testis / Q000254:growth & development / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics*; D014960:X Chromosome / Q000235:genetics; D014998:Y Chromosome / Q000235:genetics
chemical_list: D004268:DNA-Binding Proteins; D051741:Kruppel-Like Transcription Factors; D014157:Transcription Factors; C501342:Zfy1 protein, mouse; C062076:zinc finger protein, X-linked