Generation of Cas9 transgenic zebrafish and their application in establishing an ERV-deficient animal model

Zhe Yang; Shihao Chen; Songlei Xue; Xinxiu Li; Zhen Sun; Yu Yang; Xuming Hu; Tuoyu Geng; Hengmi Cui

doi:10.1007/s10529-018-2605-5

Generation of Cas9 transgenic zebrafish and their application in establishing an ERV-deficient animal model

Biotechnol Lett. 2018 Dec;40(11-12):1507-1518. doi: 10.1007/s10529-018-2605-5. Epub 2018 Sep 22.

Authors

Zhe Yang^{1

2}, Shihao Chen^{1

2}, Songlei Xue^{1

2}, Xinxiu Li^{1

2}, Zhen Sun^{1

2}, Yu Yang^{1

2}, Xuming Hu^{1

2}, Tuoyu Geng^{1

2

3}, Hengmi Cui^{4

5

6

7

8}

Affiliations

¹ Institute of Epigenetics and Epigenomics, Yangzhou University, Yangzhou, Jiangsu, 225009, China.
² College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu, 225009, China.
³ College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu, 225000, China.
⁴ Institute of Epigenetics and Epigenomics, Yangzhou University, Yangzhou, Jiangsu, 225009, China. hmcui@yzu.edu.cn.
⁵ College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu, 225009, China. hmcui@yzu.edu.cn.
⁶ Institute of Comparative Medicine, Yangzhou University, Yangzhou, Jiangsu, 225009, China. hmcui@yzu.edu.cn.
⁷ Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, Jiangsu, 225009, China. hmcui@yzu.edu.cn.
⁸ Joint International Research Laboratory of Agricultural & Agri-Product Safety, The Ministry of Education of China, Yangzhou University, Yangzhou, Jiangsu, 225009, China. hmcui@yzu.edu.cn.

Abstract

Objectives: To investigate the effect of endogenous Cas9 on genome editing efficiency in transgenic zebrafish.

Results: Here we have constructed a transgenic zebrafish strain that can be screened by pigment deficiency. Compared with the traditional CRISPR injection method, the transgenic zebrafish can improve the efficiency of genome editing significantly. At the same time, we first observed that the phenotype of vertebral malformation in early embryonic development of zebrafish after ZFERV knockout.

Conclusions: The transgenic zebrafish with expressed Cas9, is more efficient in genome editing. And the results of ZFERV knockout indicated that ERV may affect the vertebral development by Notch1/Delta D signal pathway.

Keywords: CRISPR/Cas9; Embryonic development; Genomic editing; Spinal abnormality; Zebrafish.

MeSH terms

Animals
Animals, Genetically Modified / genetics*
CRISPR-Cas Systems / genetics*
Endogenous Retroviruses / genetics*
Female
Gene Editing / methods*
Gene Knockout Techniques
Male
Promoter Regions, Genetic / genetics
Zebrafish / genetics*

Abstract

MeSH terms

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