CLC number:
On-line Access: 2022-01-12
Received: 2021-04-11
Revision Accepted: 2021-06-09
Crosschecked: 0000-00-00
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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",
volume="23",
number="1",
pages="74-83",
year="2022",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B2100343"
}
%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 Zhi WANG
%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
TY - JOUR
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
Abstract: 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.
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