PUBLICATION

A novel technique based on in vitro oocyte injection to improve CRISPR/Cas9 gene editing in zebrafish

Authors
Xie, S.L., Bian, W.P., Wang, C., Junaid, M., Zou, J.X., Pei, D.S.
ID
ZDB-PUB-160930-10
Date
2016
Source
Scientific Reports   6: 34555 (Journal)
Registered Authors
Bian, Wanping, Junaid, Muhammad, Pei, Desheng, Wang, Chao, Xie, Shaolin
Keywords
Animal breeding, Gene targeting, Mutagenesis
MeSH Terms
none
PubMed
27680290 Full text @ Sci. Rep.
Abstract
Contemporary improvements in the type II clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system offer a convenient way for genome editing in zebrafish. However, the low efficiencies of genome editing and germline transmission require a time-intensive and laborious screening work. Here, we reported a method based on in vitro oocyte storage by injecting oocytes in advance and incubating them in oocyte storage medium to significantly improve the efficiencies of genome editing and germline transmission by in vitro fertilization (IVF) in zebrafish. Compared to conventional methods, the prior micro-injection of zebrafish oocytes improved the efficiency of genome editing, especially for the sgRNAs with low targeting efficiency. Due to high throughputs, simplicity and flexible design, this novel strategy will provide an efficient alternative to increase the speed of generating heritable mutants in zebrafish by using CRISPR/Cas9 system.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping