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

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...

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Detalles Bibliográficos
Autores principales: Xiao, Guohui, Fu, Xiangdong, Zhang, Juanjuan, Liu, Shuyan, Wang, Zhaoqin, Ye, Taosheng, Zhang, Guoliang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Netherlands 2021
Materias:
Original Research Paper
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8445740/
https://www.ncbi.nlm.nih.gov/pubmed/34532823
http://dx.doi.org/10.1007/s10529-021-03177-z
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author Xiao, Guohui
Fu, Xiangdong
Zhang, Juanjuan
Liu, Shuyan
Wang, Zhaoqin
Ye, Taosheng
Zhang, Guoliang
author_facet Xiao, Guohui
Fu, Xiangdong
Zhang, Juanjuan
Liu, Shuyan
Wang, Zhaoqin
Ye, Taosheng
Zhang, Guoliang
author_sort Xiao, Guohui
collection PubMed
description 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. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10529-021-03177-z.
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spelling pubmed-84457402021-09-17 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 Biotechnol Lett Original Research Paper 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. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10529-021-03177-z. Springer Netherlands 2021-09-17 2021 /pmc/articles/PMC8445740/ /pubmed/34532823 http://dx.doi.org/10.1007/s10529-021-03177-z Text en © The Author(s), under exclusive licence to Springer Nature B.V. 2021 This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Original Research Paper
Xiao, Guohui
Fu, Xiangdong
Zhang, Juanjuan
Liu, Shuyan
Wang, Zhaoqin
Ye, Taosheng
Zhang, Guoliang
Rapid and cost-effective screening of CRISPR/Cas9-induced mutants by DNA-guided Argonaute nuclease
title Rapid and cost-effective screening of CRISPR/Cas9-induced mutants by DNA-guided Argonaute nuclease
title_full Rapid and cost-effective screening of CRISPR/Cas9-induced mutants by DNA-guided Argonaute nuclease
title_fullStr Rapid and cost-effective screening of CRISPR/Cas9-induced mutants by DNA-guided Argonaute nuclease
title_full_unstemmed Rapid and cost-effective screening of CRISPR/Cas9-induced mutants by DNA-guided Argonaute nuclease
title_short Rapid and cost-effective screening of CRISPR/Cas9-induced mutants by DNA-guided Argonaute nuclease
title_sort rapid and cost-effective screening of crispr/cas9-induced mutants by dna-guided argonaute nuclease
topic Original Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8445740/
https://www.ncbi.nlm.nih.gov/pubmed/34532823
http://dx.doi.org/10.1007/s10529-021-03177-z
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