A simple and rapid method for enzymatic synthesis of CRISPR-Cas9 sgRNA libraries.

ABSTRACT: CRISPR-Cas9 sgRNA libraries have transformed functional genetic screening and have enabled several innovative methods that rely on simultaneously targeting numerous genetic loci. Such libraries could be used in a vast number of biological systems and in the development of new technologies, but library generation is hindered by the cost, time, and sequence data required for sgRNA library synthesis. Here, we describe a rapid enzymatic method for generating robust, variant-matched libraries from any source of cDNA in under 3 h. This method, which we have named SLALOM, utilizes a custom sgRNA scaffold sequence and a novel method for detaching oligonucleotides from solid supports by a strand displacing polymerase. With this method, we constructed libraries targeting the E. coli genome and the t

SUBMITTER: Yates JD 

PROVIDER: S-EPMC8682767 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature