PubMed 29872452

PubMed ID: 29872452

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A simple and cost-effective method for screening of CRISPR/Cas9-induced homozygous/biallelic mutants.
Authors: Guo Jinggong, Li Kun, Jin Lifeng, Xu Rui, Miao Kaiting, Yang Fengbo, Qi Chaoya, Zhang Lin, Botella Jose R, Wang Ran, Miao Yuchen
Journal: Plant methods (Plant Methods), Vol.14(), 2018

DOI: 10.1186/s13059-015-0715-0 PMCID: PMC4507317

Abstract
The CRISPR/Cas9 system is being used for genome editing purposes by many research groups in multiple plant species. Traditional sequencing methods to identify homozygous mutants are time-consuming, laborious and expensive.

We have developed a method to screen CRISPR/Cas9-induced mutants through Mutation Sites Based Specific Primers Polymerase Chain Reaction (MSBSP-PCR). The MSBSP-PCR method was successfully used to identify homozygous/biallelic mutants in Nicotiana tabacum and Arabidopsis thaliana, and we speculate that it can be used for the identification of CRISPR/Cas9-induced mutants in other plant species. Compared to traditional sequencing methods, MSBSP-PCR is simpler, faster and cheaper.

The MSBSP-PCR method is simple to implement and can save time and cost in the screening of CRISPR/Cas9-induced homozygous/biallelic mutants.
Publication Types
Journal Article
Keywords
Arabidopsis thaliana CRISPR/Cas9 Genome editing Mutant screening PCR Tobacco
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