|ZFIN ID: ZDB-PUB-160226-17|
A comparison of the knockout efficiencies of two codon-optimized Cas9 coding sequences in zebrafish embryos
Fenghua, Z., Houpeng, W., Siyu, H., Feng, X., Zuoyan, Z., Yonghua, S.
|Source:||Yi chuan = Hereditas 38: 144-54 (Journal)|
|Registered Authors:||Zhu, Zuoyan|
|Keywords:||CRISPR/Cas9, zebrafish, gene knockout, mutation efficiency|
|PubMed:||26907778 Full text @ Yi Chuan|
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.
ABSTRACTRecent 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.
ADDITIONAL INFORMATION No data available
ERRATA and NOTESArticle in Chinese.