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On-line Access: 2022-01-12

Received: 2021-04-11

Revision Accepted: 2021-06-09

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Tiansheng CHEN


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Journal of Zhejiang University SCIENCE B 2022 Vol.23 No.1 P.74-83


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

Author(s):  Qihua PAN, Junzhi LUO, Yuewen JIANG, Zhi WANG, Ke LU, Tiansheng CHEN

Affiliation(s):  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; more

Corresponding email(s):   tiansheng.chen@jmu.edu.cn

Key Words:  Medaka (Oryzias latipes), Gene editing, Poly-tRNA-gRNA, Embryos, Fish cells

Qihua PAN, Junzhi LUO, Yuewen JIANG, Zhi WANG, Ke LU, Tiansheng CHEN. Efficient gene editing in a medaka (Oryzias latipes) cell line and embryos by SpCas9/tRNA-gRNA[J]. Journal of Zhejiang University Science B, 2022, 23(1): 74-83.

@article{title="Efficient gene editing in a medaka (Oryzias latipes) cell line and embryos by SpCas9/tRNA-gRNA",
author="Qihua PAN, Junzhi LUO, Yuewen JIANG, Zhi WANG, Ke LU, Tiansheng CHEN",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Efficient gene editing in a medaka (Oryzias latipes) cell line and embryos by SpCas9/tRNA-gRNA
%A Qihua PAN
%A Junzhi LUO
%A Yuewen JIANG
%A Ke LU
%A Tiansheng CHEN
%J Journal of Zhejiang University SCIENCE B
%V 23
%N 1
%P 74-83
%@ 1673-1581
%D 2022
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B2100343

T1 - Efficient gene editing in a medaka (Oryzias latipes) cell line and embryos by SpCas9/tRNA-gRNA
A1 - Qihua PAN
A1 - Junzhi LUO
A1 - Yuewen JIANG
A1 - Zhi WANG
A1 - Ke LU
A1 - Tiansheng CHEN
J0 - Journal of Zhejiang University Science B
VL - 23
IS - 1
SP - 74
EP - 83
%@ 1673-1581
Y1 - 2022
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B2100343

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.

利用化脓链球菌成簇的规则间隔短回文重复序列相关蛋白9(spCas9)/转运RNA(tRNA)-导向RNA(gRNA)(SpCas9/tRNA-gRNA)在青鳉(Oryzias latipes)细胞系和胚胎中进行高效基因编辑


关键词:青鳉(Oryzias latipes);基因编辑;Poly-tRNA-gRNA;胚胎;鱼类细胞系

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


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