利用CRISPR/Cas9技术进行Adcy3基因编辑小鼠模型的构建与鉴定

Construction and Confirmation of Adcy3 Gene Editing Mouse Model by CRISPR/Cas9 Technology

目的:利用CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR- As-sociated 9)技术构建成功Adcy3 (Cyclic Adenosine-3',5'-Monophosphate, cAMP)下游插入mCherry基因小鼠品系,为深入研究该基因的功能及相关疾病的治疗提供条件。方法:设计Adcy3基因sgRNA和mCherry供体,通过显微注射方式将Cas9mRNA、sgRNA和mCherry供体一同注射到C57BL/6小鼠受精卵中,获得Adcy3基因编辑小鼠。经过pcr鉴定和Sanger测序,对获得的子代小鼠,进行基因型的鉴定。结果:Adcy3基因编辑鼠构建成功,且能够通过正常繁育得到能够稳定遗传的子代小鼠。结论:建立了Adcy3-基因编辑小鼠模型,为小鼠体内Adcy3基因功能研究提供了新的小鼠模型。

Objective: A mouse strain with mCherry gene inserted downstream of Adcy3 was successfully con-structed by CRISPR/Cas9 Technology, which provided a convenient animal models for in-depth study of the function of this gene and treatment of related diseases. Methods: Adcy3 gene sgRNA and mCherry donors were designed. Cas9 mRNA, sgRNA and mCherry donors were microinjected into the fertilized eggs of C57BL/6 mice to obtain Adcy3 gene editing mice. After pcr identification and Sanger sequencing, the obtained offspring mice were genotyped. Results: Adcy3 gene editing mice were successfully constructed, and could obtain stable offspring mice through normal breed-ing. Conclusions: It was confirmed that the Adcy3-mCherry mouse line was generated successfully, which provided a new mouse model for future studies of Adcy3 function research in vivo.