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Targeted mutagenesis in the olive flounder (Paralichthys olivaceus) using the CRISPR/Cas9 system with electroporation

文献类型：期刊论文

作者Wang, Ling3,4,5; Tan, Xungang3,4; Wu, Zhihao3,4; Wang, Lijuan3,4; Jiao, Shuang3,4; Zou, Yuxia3,4; Ji, Guanglei1,2; You, Feng3,4
刊名BIOLOGIA
出版日期2021-01-20
页码8
关键词Genome editing CRISPR/Cas9 system Electroporation Myomaker and gsdf Olive flounder (Paralichthys olivaceus)
ISSN号0006-3088
DOI10.2478/s11756-020-00677-7
通讯作者Tan, Xungang(tanx@qdio.ac.cn) ; You, Feng(youfeng@qdio.ac.cn)
英文摘要As a new breeding technology, genome editing becomes a powerful tool owing to its high efficiency of gene targeting. In CRISPR/Cas9 system, how to efficiently transfer gRNA and Cas9 mRNA into embryos is an important step. Though microinjection is the most common method for operating on fish embryos, it is not easy to inject the RNA into pelagic and telolecithal eggs with hard egg chorion, such as the olive flounder (Paralichthys olivaceus) eggs. Therefore, an efficient and simple technology is urgently needed for this kind of study. In the present study, we used the electroporation method to introduce foreign gene into the flounder eggs. The results showed that the proper electroporation condition was 3 pulses for 1 millisecond (ms), 50 ms interval, at 25 V with high survival rate. Under this condition, the effect of CRISPR/Cas9 system on genome editing by using two different genes, myomaker and gonadal soma derived factor (gsdf) was investigated. Around 12% and 7% of the electroporated embryos for myomaker and gsdf hatched, respectively. The mutation sites including insert and deletion mutations at the candidate sites were visible for both targeted genes in the hatched larvae. The checked frame-shift and start codon deletion mutations would lead to complete destruction of these genes' structure. Above results implied that CRISPR/Cas9 system could work well in marine fish with pelagic eggs by using electroporation, and genome editing could be achieved on a large scale which may be useful for study of gene function in marine fish.
资助项目National Key R&D Program of China[2018YFD0900202] ; National Sciences of Foundation of China[31672636] ; National Sciences of Foundation of China[31772834]
WOS研究方向Life Sciences & Biomedicine - Other Topics
语种英语
WOS记录号WOS:000609070200004
出版者SPRINGER
源URL[http://ir.qdio.ac.cn/handle/337002/170063]  
专题海洋研究所_实验海洋生物学重点实验室
通讯作者Tan, Xungang; You, Feng
作者单位1.Weihai Fisheries Technol Promot Stn, Weihai, Peoples R China
2.Shenghang Aquat Sci & Technol Co Ltd, Weihai, Peoples R China
3.Chinese Acad Sci, Ctr Ocean Megasci, Inst Oceanol, CAS Key Lab Expt Marine Biol, 7 Nanhai Rd, Qingdao 266071, Peoples R China
4.Pilot Natl Lab Marine Sci & Technol Qingdao, Lab Marine Biol & Biotechnol, Qingdao 266237, Peoples R China
5.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
推荐引用方式
GB/T 7714		 Wang, Ling,Tan, Xungang,Wu, Zhihao,et al. Targeted mutagenesis in the olive flounder (Paralichthys olivaceus) using the CRISPR/Cas9 system with electroporation[J]. BIOLOGIA,2021:8.
APA Wang, Ling.,Tan, Xungang.,Wu, Zhihao.,Wang, Lijuan.,Jiao, Shuang.,...&You, Feng.(2021).Targeted mutagenesis in the olive flounder (Paralichthys olivaceus) using the CRISPR/Cas9 system with electroporation.BIOLOGIA,8.
MLA Wang, Ling,et al."Targeted mutagenesis in the olive flounder (Paralichthys olivaceus) using the CRISPR/Cas9 system with electroporation".BIOLOGIA (2021):8.

入库方式： OAI收割

来源：海洋研究所

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