Knowledge Network Node
Hereditas. 2016, (12) Core

Establishment of porcine Xist knockout model using CRISPR/Cas9 systemChinese Full Text

Guoling Li;Cuili Zhong;Sheng Ni;Dewu Liu;Gengyuan Cai;Zicong Li;Huaqiang Yang;Zhenfang Wu;National Engineering Research Center for Swine Breeding Industry, College of Animal Science,South China Agricultural University;

Abstract: Somatic cell nuclear transfer technique has great applications in livestock breeding, production of genetically modified animals, rescue of endangered species and treatment of human diseases. However, the currently low efficiency in animals cloning, an average of less than 5%, greatly hindered the rapid development of this tec hnique. Among many factors which affect the efficiency of cloning pigs, X chromosome inactivation is an important one. Moreover, Xist gene is closely related to X chromosome inactivation, suggesting that it may directly or indirectly affects cloning efficiency. In this study, multiple sg RNAs were designed based on the CRISPR/Cas system, and two sites（Target 3 and Target 4） whose mutation efficiency were 1% and 3% at the cellular lev el were selected. We successfully knocked out Xist with 100% efficiency by microinjecting sg RNAs for Target 3 and Target 4 in embryo. Finally, 6 cloning piglets were born including two Xist-fully-knockout piglets. The follow-up studies on increasing cloning efficiency can be carried out based on the Xist-knockout model.
Keywords：

Xist; CRISPR/Cas; cloning efficiency; porcine;

  • DOI：

    10.16288/j.yczz.16-137

  • Series：

  • Subject：

  • Classification Code：

    Q78

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