一种电转染华贵栉孔扇贝基因编辑的应用方法  

An application method for gene editing mediated by electroporation in the noble scallop Chlamys nobilis

作　　者：陈颖 张洪宽 何成 Tan Karsoon 郑怀平[1,2,3] CHEN Ying;ZHANG Hongkuan;HE Cheng;TAN Karsoon;ZHENG Huaiping(Marine Sciences Institute,Shantou University,Shantou 515063,China;Key Laboratory of Marine Biotechnology of Guangdong Province,Shantou 515063,China;Research Center for Subtropical Mariculture of Guangdong Province,Shantou 515063,China)

机构地区：[1]汕头大学海洋科学研究院,广东汕头515063 [2]广东省海洋生物技术重点实验室,广东汕头515063 [3]广东省亚热带海水养殖工程技术研究中心,广东汕头515063

出　　处：《海洋通报》2023年第1期66-73,共8页Marine Science Bulletin

基　　金：国家重点研发计划(2018YFD0901400);广东省重点领域研发计划(2021B0202020003)。

摘　　要：目前,CRISPR/Cas9基因编辑技术已成为动植物遗传育种领域研究的热点。为解决CRISPR技术在贝类生产实践中应用难的问题,本文探究了一种电转染的双壳贝类基因编辑方法。利用本方法分别对华贵栉孔扇贝受精卵的SRB-like-3和MSTN进行编辑,并成功地将编辑后的幼虫培养到稚贝。在编辑后5 d和10 d的D-型幼虫及40 d的稚贝中都检测到了荧光,并且编辑MSTN后的10 d幼虫,其壳长显著大于编辑SRB-like-3组和对照组。本研究表明:电转染基因编辑技术在贝类中具有可行性。本方法具有成本低、操作简单、易推广等优点,可以广泛应用于贝类的遗传育种领域。At present,the application of CRISPR/Cas9 gene editing technique has become a research hotspot in the field of genetics and breeding in animals and plants.In order to solve the problem in mollusks,an application method for CRISPR gene editing technique mediated by electroporation in bivalves was developed in the present study.Two genes of SRB-like-3(a key gene related to carotenoids accumulation)and MSTN(myostatin gene)were both successfully edited in the fertilized eggs of the noble scallop Chlamys nobilis using the method.And,the fluorescence was detected in D-larvae at 5 and 10 days,as well as juveniles at 40 days after the two genes edited,respectively.Moreover,shell length of D-larvae at 10 days from editing MSTN group was both significantly larger(P<0.05)than that from editing SRB-like-3 group and control group.This study shows that the application method for CRISPR/Cas9 gene editing mediated by electroporation in bivalves is feasible.More importantly,the method has some advantages of low cost,simple operation and easy to popularize,etc.,so it will be widely applied to the filed of genetics and breeding in mollusks.

关 键 词：华贵栉孔扇贝 CRISPR/Cas9 电转染 基因编辑 

分 类 号：P736.22[天文地球—海洋地质]