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

摘要:
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。

Abstract:
ObjectiveTo construct a Trib3 knockout rat model and identify the related genotype.
MethodsAfter 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.
ResultsThe 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.
ConclusionsTrib3 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.