Design of a generic CRISPR-Cas9 approach using the same sgRNA to perform gene editing at distinct loci.

ABSTRACT: BACKGROUND:The CRISPR/Cas (clustered regularly interspaced short palindromic repeat and CRISPR-associated nucleases) based technologies have revolutionized genome engineering. While their use for prokaryotic genome editing is expanding, some limitations remain such as possible off-target effects and design constraints. These are compounded when performing systematic genome editing at distinct loci or when targeting repeated sequences (e.g. multicopy genes or mobile genetic elements). To overcome these limitations, we designed an approach using the same sgRNA and CRISPR-Cas9 system to independently perform gene editing at different loci. RESULTS:We developed a two-step procedure based on the introduction by homologous recombination of 'bait' DNA at the vicinity of a gene copy of interest be

SUBMITTER: Najah S 

PROVIDER: S-EPMC6425556 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature