Full Text:
<35>
CLC number:
On-line Access: 2024-04-24
Received: 2023-12-11
Revision Accepted: 2024-03-26
Crosschecked: 0000-00-00
Cited: 0
Clicked: 49
Efficient genome editing in Medaka (Oryzias latipes) using a codon optimized SaCas9 system
| |||||||||||||
Yuewen JIANG, Qihua PAN, Zhi WANG, Ke LU, Bilin XIA, Tiansheng CHEN. Efficient genome editing in Medaka (Oryzias latipes) using a codon optimized SaCas9 system[J]. Journal of Zhejiang University Science B, 1998, -1(-1): .
@article{title="Efficient genome editing in Medaka (Oryzias latipes) using a codon optimized SaCas9 system",
author="Yuewen JIANG, Qihua PAN, Zhi WANG, Ke LU, Bilin XIA, Tiansheng CHEN",
journal="Journal of Zhejiang University Science B",
volume="-1",
number="-1",
pages="",
year="1998",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B2300899"
}
%0 Journal Article
%T Efficient genome editing in Medaka (Oryzias latipes) using a codon optimized SaCas9 system
%A Yuewen JIANG
%A Qihua PAN
%A Zhi WANG
%A Ke LU
%A Bilin XIA
%A Tiansheng CHEN
%J Journal of Zhejiang University SCIENCE B
%V -1
%N -1
%P
%@ 1673-1581
%D 1998
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B2300899
TY - JOUR
T1 - Efficient genome editing in Medaka (Oryzias latipes) using a codon optimized SaCas9 system
A1 - Yuewen JIANG
A1 - Qihua PAN
A1 - Zhi WANG
A1 - Ke LU
A1 - Bilin XIA
A1 - Tiansheng CHEN
J0 - Journal of Zhejiang University Science B
VL - -1
IS - -1
SP -
EP -
%@ 1673-1581
Y1 - 1998
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B2300899
Abstract: The CRISPR-Cas9 system, belonging to the type II CRISPR/Cas system, is an effective gene editing tool widely used in different organisms, but the size of SpCas9 is quite large (4.3 kb), which is not convenient for vector delivery. In this study, we used a codon-optimized saCas9 system to edit the tyr, oca2, and pax6.1 genes in the fish model medaka (Oryzias latipes), in which the size of saCas9 (3.3 kb) is much smaller and the necessary prototype spacer-adjacent motif (PAM) sequence is 5'-NNGRRT-3'. We also used a tRNA-sgRNA system to express the functional sgRNA by transcription either in vivo or in vitro, and the combination of saCas9 and tRNA-sgRNA was used to edit the tyr gene in the medaka genome. The saCas9/sgRNA and saCas9/tRNA-sgRNA systems were shown to edit the medaka genome effectively, while the PAM sequence is an essential part for the efficiency of editing. Besides, tRNA can improve the flexibility of the system by enabling the sgRNA to be controlled by a common promoter such as cytomegalovirus. Moreover, the all-in-one cassette CMV-saCas9-tRNA-sgRNA-tRNA is functional in medaka gene editing. Taken together, the codon-optimized saCas9 system provides an alternative and smaller tool to edit the medaka genome and potentially other fish genomes.
Open peer comments: Debate/Discuss/Question/Opinion
<1>