一种电转染华贵栉孔扇贝基因编辑的应用方法

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。

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

At present, the application of gene editing technique of CRISPR/Cas9 has become 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 gene editing of CRISPR/Cas9 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. The present study showed that the application method for gene editing of CRISPR/Cas9 mediated by electroporation in bivalves was feasible. More importantly, the method has some advantages of low cost, simple operation and easy to popularize, etc., so it will be widely applied in the filed of genetics and breeding in mollusks.