应用CRISPR/Cas9技术敲除Trib3基因大鼠的基因型鉴定

Genotype identification of Trib3 knockout rats using CRISPR/Cas9 technology

目的:构建Trib3基因敲除大鼠模型及基因型鉴定。方法:靶向敲除大鼠Trib3基因后,通过基因测序和PCR技术对大鼠Trib3基因的第3号外显子删除情况进行检测与基因型鉴定。结果:得到阳性F0代首建大鼠Trib3^(-/+),通过自交得到F1与F2代大鼠,筛选出F2代Trib3^(-/-)大鼠。结论:成功制备Trib3基因敲除大鼠模型,依次鉴定出Trib3^(-/-)、Trib3^(-/+)和Trib3^(+/+)大鼠,并可以稳定遗传,为后续的实验开展提供理论依据。

Objective:To construct a Trib3 knockout rat model and identify the related genotype.Methods:After targeted knockout of Trib3 gene in rat model,the deletion of exon 3 of Trib3 gene was detected,and the genotype was identified by gene sequencing and PCR methods.Results:The positive F0 generation of Trib3^(-/+)rats was obtained,the F1 and F2 generations were obtained by self-crossing,and the F2 generation of Trib3^(-/-)rats was selected.Conclusions:Trib3 knockout rat models are successfully prepared,and Trib3^(-/-),Trib3^(-/+)and Trib3^(+/+)rats are successively identified and can be stably inherited,it provides a theoretical basis for subsequent experiments.