PUBLICATION
A comparison of the knockout efficiencies of two codon-optimized Cas9 coding sequences in zebrafish embryos
- Authors
- Fenghua, Z., Houpeng, W., Siyu, H., Feng, X., Zuoyan, Z., Yonghua, S.
- ID
- ZDB-PUB-160226-17
- Date
- 2016
- Source
- Yi chuan = Hereditas 38: 144-54 (Journal)
- Registered Authors
- Zhu, Zuoyan
- Keywords
- CRISPR/Cas9, zebrafish, gene knockout, mutation efficiency
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- Animals, Genetically Modified
- Base Sequence
- CRISPR-Cas Systems*
- Codon/genetics*
- Embryo, Nonmammalian/embryology
- Embryo, Nonmammalian/metabolism*
- Female
- Gene Expression Regulation, Developmental
- Green Fluorescent Proteins/genetics
- Green Fluorescent Proteins/metabolism
- Male
- Molecular Sequence Data
- RNA, Messenger/genetics
- RNA, Messenger/metabolism
- Sequence Homology, Amino Acid
- Time Factors
- Zebrafish/embryology
- Zebrafish/genetics*
- Zebrafish Proteins/genetics*
- Zebrafish Proteins/metabolism
- PubMed
- 26907778 Full text @ Yi Chuan
Citation
Fenghua, Z., Houpeng, W., Siyu, H., Feng, X., Zuoyan, Z., Yonghua, S. (2016) A comparison of the knockout efficiencies of two codon-optimized Cas9 coding sequences in zebrafish embryos. Yi chuan = Hereditas. 38:144-54.
Abstract
Recent years have witnessed the rapid development of the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein(CRISPR/Cas9)system. In order to realize gene knockout with high efficiency and specificity in zebrafish, several labs have synthesized distinct Cas9 cDNA sequences which were cloned into different vectors. In this study, we chose two commonly used zebrafish-codon-optimized Cas9 coding sequences (zCas9_bz, zCas9_wc) from two different labs, and utilized them to knockout seven genes in zebrafish embryos, including the exogenous egfp and six endogenous genes (chd, hbegfa, th, eef1a1b, tyr and tcf7l1a). We compared the knockout efficiencies resulting from the two zCas9 coding sequences, by direct sequencing of PCR products, colony sequencing and phenotypic analysis. The results showed that the knockout efficiency of zCas9_wc was higher than that of zCas9_bz in all conditions.
Errata / Notes
Article in Chinese.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping