靶向小鼠CREPT基因CRISPR/Cas9敲除质粒的构建  

Construction of CRISPR/Cas9 Gene Knockout Plasmids Targeting Mouse CREPT Gene

作　　者：刘素丽 范柠粼 邹晓辉 武会娟 LIU Suli;FAN Ninglin;ZOU Xiaohui;WU Huijuan(Beijing Laboratory Animal Research Center Co.,Ltd,Beijing 102609,China;China-Janpan Friendship Hospital,Beijing 100029,China)

机构地区：[1]北京实验动物研究中心有限公司,北京102609 [2]中日友好医院,北京100029

出　　处：《实验动物科学》2023年第1期79-82,共4页Laboratory Animal Science

摘　　要：目的利用规律间隔成簇短回文重复序列/相关蛋白9(CRISPR/Cas9)系统构建靶向小鼠CREPT基因的敲除质粒。方法根据CRISPR/Cas9靶点设计原则靶向小鼠CREPT基因的sgRNAs,与体外转录载体pUC57-sgRNA连接,构建CREPT基因敲除质粒;体外切割实验验证设计的sgRNAs是否具有活性。结果设计了3条sgRNAs并分别准确插入pUC57-sgRNA载体;体外切割实验结果显示,以上3条sgRNAs均能够介导Cas9蛋白对靶片断进行切割。结论制备了小鼠CREPT基因敲除质粒和体外转录的sgRNAs,为构建CREPT基因敲除小鼠模型奠定了重要基础。Objective To construct CRISPR/Cas9 gene knockout plasmids targeting mouse CREPT gene.Method sgRNAs targeting mouse CREPT gene were designed according to the CRISPR/Cas9 target design principles and were cloned into the in vitro transcription vector pUC57-sgRNA to construct CREPT gene knockout plasmids.Result Three sgRNAs were designed and cloned into puc57-sgRNA,in vitro digestion experiment showed that the above three sgRNAs could mediate Cas9 protein to cleave the target fragments.Conclusion Mouse CREPT gene knockout plasmids and in vitro transcribed sgRNAs were prepared,laying an important foundation for construction of gene knockout mouse model.

关 键 词：CREPT CRISPR/Cas9 基因敲除 

分 类 号：Q95-3[生物学—动物学]