利用CRISPR/Cas9技术进行lncRNA snhg17-KO基因敲除小鼠模型的构建与鉴定

LncRNA snhg17-KO Gene Knockout Mouse Model was Constructed and Identified Using CRISPR/Cas9 Technique

目的:构建lncRNA snhg17-KO基因敲除小鼠模型,为探讨snhg17在肿瘤发生中的作用机制及体内实验研究提供更特异可靠的动物模型。方法:确定snhg17-KO基因敲除小鼠特异性靶标sgRNA1、sgRNA2,引导与核酸酶Cas9结合进行RNA的体外转录,将有活性的sgRNA和Cas9RNA通过C57BL/6小鼠受精卵的显微注射方式,获取snhg17基因敲除的纯合子小鼠。运用聚合酶链反应(PCR)和基因测序方法,针对获得的子代小鼠,进行基因型的鉴定。结果:lncRNA snhg17-KO基因敲除小鼠模型构建成功。结论:本研究采用CRISPR/Cas9技术首次构建出snhg17基因敲除小鼠的动物模型,为探讨snhg17在肿瘤发生、发展中的作用提供更为便捷、可靠的动物模型及研究平台。

Objective:LncRNA snhg17-KO gene knockout mouse model was constructed.To provide a more specific and reliable animal model to explore the mechanism of snhg17 in tumorigenesis and in vivo experimental study.Methods:The specific targets of snhg17-KO gene knockout mice,sgRNA1 and sgRNA2,were determined.RNA was transcribed with Cas9 nuclease in vitro.The active sgRNA and Cas9RNA were microinjected into the fertilized eggs of C57BL/6 mice,and the snhg17 knockout homozygous mice were obtained.The genotypes of the offspring mice were identified by PCR polymerase chain reaction and gene sequencing.Results:LncRNA snhg17-KO gene knockout mouse model was successfully constructed.Conclusion:Using CRISPR/Cas9 gene knockout technique,a mouse model of snhg17 gene knockout was established for the first time.To explore the role of snhg17 in tumor genesis and development,and to provide convenient and reliable animal models and research platforms.