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 simp

SUBMITTER: Guo J 

PROVIDER: S-EPMC5972395 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature