Improvement of developmental competence of cloned male pig embryos by short hairpin ribonucleic acid (shRNA) vector-based but not small interfering RNA (siRNA)-mediated RNA interference (RNAi) of Xist expression
文献类型： 外文期刊
作者： Yang, Xuqiong 1 ; Wu, Xiao 1 ; Yang, Yang 1 ; Gu, Ting 1 ; Hong, Linjun 1 ; Zheng, Enqin 1 ; Xu, Zheng 1 ; Zeng, Fang 3 ; Sh 1 ;
作者机构： 1.South China Agr Univ, Coll Anim Sci, Natl Engn Res Ctr Breeding Swine Ind, Guangzhou 510642, Guangdong, Peoples R China
2.South China Agr Univ, Coll Anim Sci, Guangdong Prov Key Lab Agroanim Genom & Mol Breed, Guangzhou 510642, Guangdong, Peoples R China
3.South China Agr Univ, Coll Marine Sci, Guangzhou 510642, Guangdong, Peoples R China
4.Wens Foodstuff Grp Co Ltd, Guangdong Wens Pig Breeding Technol Co Ltd, Yunfu 527400, Guangdong, Peoples R China
关键词： Cloned; Pig; Ribonucleic acid interference (RNAi); Somatic cell nuclear transfer (SCNT); Xist
期刊名称：JOURNAL OF REPRODUCTION AND DEVELOPMENT （ 影响因子：2.214； 五年影响因子：2.221 ）
ISSN： 0916-8818
年卷期： 2019 年 65 卷 6 期
页码：
收录情况： SCI
摘要： Xist is an X-linked ribonucleic acid (RNA) gene responsible for the cis induction of X chromosome inactivation (XCI). In cloned mammalian embryos, Xist is ectopically activated at the morula to blastocyst stage on the X chromosome that is supposed to be active, thus resulting in abnormal XCI. Suppression of erroneous Xist expression by injecting small interfering RNA (siRNA) remarkably increased the developmental efficiency of cloned male mouse embryos by approximately 10-fold. However, injection of anti-Xist siRNA resulted in only a slight increase in the developmental ability of injected cloned male pig embryos because the blocking effect of the injected siRNA was not maintained beyond the morula stage, which is 5 days post-activation. To develop a more effective approach for suppressing the ectopic expression of Xist in cloned pig embryos, we compared the silencing effect of short hairpin RNA (shRNA) and siRNA on Xist expression and the effects of these two Xist knockdown methods on the developmental competence of cloned male pig embryos. Results indicated that an shRNA-based RNA interference (RNAi) has a longer blocking effect on Xist expression than an siRNA-mediated RNAi. Injection of anti-Xist shRNA plasmid into two-cell-stage cloned male pig embryos effectively suppressed Xist expression, rescued XCI at the blastocyst stage, and improved the in vitro developmental ability of injected cloned embryos. These positive effects, however, were not observed in cloned male pig embryos injected with anti-Xist siRNA. This study demonstrates that vector-based rather than siRNA-mediated RNAi of Xist expression can be employed to improve pig cloning efficiency.
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