Rapid and cost-effective screening of CRISPR/Cas9-induced mutants by DNA-guided Argonaute nuclease

Xiao, Guohui; Fu, Xiangdong; Zhang, Juanjuan; Liu, Shuyan; Wang, Zhaoqin; Ye, Taosheng; Zhang, Guoliang

2021-09-01

10.1007/s10529-021-03177-z

BIOTECHNOLOGY LETTERS   影响因子和分区

0141-5492

1573-6776

Objective With the widespread application of CRISPR/Cas9 gene editing technology, new methods are needed to screen mutants quickly and effectively. Here, we aimed to develop a simple and cost-effective method to screen CRISPR/Cas9-induced mutants. Result We report a novel method to identify CRISPR/Cas9-induced mutants through a DNA-guided Argonaute nuclease derived from the archaeon Pyrococcus furiosus. We demonstrated that the Pyrococcus furiosus Argonaute (PfAgo)-based method could distinguish among biallelic mutants, monoallelic mutants and wild type (WT). Furthermore, this method was able to identify 1 bp indel mutations. Conclusion The PfAgo-based method is simple to implement and can be applied to screen biallelic mutants and mosaic mutants generated by CRISPR-Cas9 or other kinds of gene editing tools.

CRISPR Cas9 Biallelic mutants Monoallelic mutations Argonaute nuclease Pyrococcus furiosus

SCI ; EI

英语

第一 ; 通讯

National Key Research and Development Plan["2020YFA0907200","2019YFC0840602"] ; Guangdong Foundation for Basic and Applied Basic Research[2019B1515120041] ; Shenzhen Scientific and Technological Foundation["KCXFZ202002011007083","JCYJ20180228162336873"] ; Sanming Project of Medicine in Shenzhen[SZSM201911009]

Biotechnology & Applied Microbiology

Biotechnology & Applied Microbiology

WOS:000696417200001

SPRINGER

20213810912882

Genes

Biological Materials and Tissue Engineering:461.2 ; Industrial Economics:911.2

BIOLOGY & BIOCHEMISTRY

Web of Science

Southern Univ Sci & Technol, Shenzhen Peoples Hosp 3, Natl Clin Res Ctr Infect Dis, Shenzhen, Peoples R China

Xiao, Guohui,Fu, Xiangdong,Zhang, Juanjuan,et al. Rapid and cost-effective screening of CRISPR/Cas9-induced mutants by DNA-guided Argonaute nuclease[J]. BIOTECHNOLOGY LETTERS,2021,43:2105-2110.

Xiao, Guohui.,Fu, Xiangdong.,Zhang, Juanjuan.,Liu, Shuyan.,Wang, Zhaoqin.,...&Zhang, Guoliang.(2021).Rapid and cost-effective screening of CRISPR/Cas9-induced mutants by DNA-guided Argonaute nuclease.BIOTECHNOLOGY LETTERS,43,2105-2110.

Xiao, Guohui,et al."Rapid and cost-effective screening of CRISPR/Cas9-induced mutants by DNA-guided Argonaute nuclease".BIOTECHNOLOGY LETTERS 43(2021):2105-2110.