CRISPR-finder: A high throughput and cost-effective method to identify successfully edited Arabidopsis thaliana individuals.

ABSTRACT: Genome editing with the CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR associated protein) system allows mutagenesis of a targeted region of the genome using a Cas endonuclease and an artificial guide RNA. Both because of variable efficiency with which such mutations arise and because the repair process produces a spectrum of mutations, one needs to ascertain the genome sequence at the targeted locus for many individuals that have been subjected to mutagenesis. We provide a complete protocol for the generation of amplicons up until the identification of the exact mutations in the targeted region. CRISPR-finder can be used to process thousands of individuals in a single sequencing run. We successfully identified an ISOCHORISMATE SYNTHASE 1 mutant line in

SUBMITTER: Symeonidi E 

PROVIDER: S-EPMC10095899 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature