The RNAi vecter construction and verification of the functions in sexual control of Cervus elaphus Zfx gene

Abstract Background: Zinc finger protein X-linked (Zfx) was regarded to be a sex determination factor, and plays a critical role in spermatogenesis. RNAi is an effective method of silencing Zfx/Zfy mRNA expression. However, there has been little research on the use of RNAi technology to control the sex of the offspring of Cervus elaphus. The objective of this study was first to explore an efficient method to alter the Cervus elaphus offspring sex-ratio by silencing the genes Zfx during spermatogenesis.Results: Three recombinant expression vectors pLL3.7/A, pLL3.7/B and pLL3.7/C were constructed to interrupt the Zfx gene. The results showed that the expression of Zfx mRNA was significantly silenced by pLL3.7/A (P < 0.01), compared with the control group. The group injected with pLL3.7/A produced 94 Cervus elaphus, including 68 males and 26 females. The male rates (72.34%) were significantly higher than the control groups (P < 0.01).Conclusions: Our experiment suggest that the Zfx gene plays a significant role in the process of X-sperm formation. Zfx siRNA may be a useful approach to control offspring sex in Cervus elaphus.