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A simple and cost-effective method for screening of CRISPR/Cas9-induced homozygous/biallelic mutants.

ABSTRACT: Background: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. Results: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. Conclusions:The MSBSP-PCR method is simple to implement and can save time and cost in the screening of CRISPR/Cas9-induced homozygous/biallelic mutants.

SUBMITTER: Guo J

PROVIDER: S-EPMC5972395 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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A simple and cost-effective method for screening of CRISPR/Cas9-induced homozygous/biallelic mutants.

Guo Jinggong J Li Kun K Jin Lifeng L Xu Rui R Miao Kaiting K Yang Fengbo F Qi Chaoya C Zhang Lin L Botella Jose R JR Wang Ran R Miao Yuchen Y

Plant methods 20180529

<h4>Background</h4>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.<h4>Results</h4>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 <i>Nic ...[more]

PMID: 29872452

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