Efficient gene editing in a medaka(Oryzias latipes)cell line and embryos by SpCas9/tRNA-gRNA  

作　　者：Qihua PAN Junzhi LUO Yuewen JIANG Zhi WANG Ke LU Tiansheng CHEN 

机构地区：[1]Key Laboratory of Healthy Mariculture for the East China Sea,Ministry of Agriculture and Rural Affairs,Engineering Research Center of the Modern Technology for Eel Industry,Ministry of Education,Fisheries College,Jimei University,Xiamen,361021,China [2]College of Fisheries,Key Laboratory of Freshwater Animal Breeding,Ministry of Agriculture and Rural Affairs,Huazhong Agricultural University,Wuhan,430070,China

出　　处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2022年第1期74-83,共10页浙江大学学报（英文版）B辑（生物医学与生物技术）

基　　金：This study was supported by the National Natural Science Foundation of China(Nos.31771648 and 31672653);the Scientific Research Foundation of Jimei University(No.ZQ2020003);the National Key Basic Research Program of China(No.2013CB967700).

摘　　要：Generation of mutants with clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)is commonly carried out in fish species by co-injecting a mixture of Cas9 messenger RNA(mRNA)or protein and transcribed guide RNA(gRNA).However,the appropriate expression system to produce functional gRNAs in fish embryos and cells is rarely present.In this study,we employed a poly-transfer RNA(tRNA)-gRNA(PTG)system driven by cytomegalovirus(CMV)promoter to target the medaka(Oryzias latipes)endogenous gene tyrosinase(tyr)or paired box 6.1(pax6.1)and illustrated its function in a medaka cell line and embryos.The PTG system was combined with the CRISPR/Cas9 system under high levels of promoter to successfully induce gene editing in medaka.This is a valuable step forward in potential application of the CRISPR/Cas9 system in medaka and other teleosts.

关 键 词：Medaka(Oryzias latipes) Gene editing Poly-tRNA-gRNA Embryos Fish cells 

分 类 号：S943[农业科学—水产养殖]