CLUE: a bioinformatic and wet-lab pipeline for multiplexed cloning of custom sgRNA libraries.

ABSTRACT: The systematic perturbation of genomes using CRISPR/Cas9 deciphers gene function at an unprecedented rate, depth and ease. Commercially available sgRNA libraries typically contain tens of thousands of pre-defined constructs, resulting in a complexity challenging to handle. In contrast, custom sgRNA libraries comprise gene sets of self-defined content and size, facilitating experiments under complex conditions such as in vivo systems. To streamline and upscale cloning of custom libraries, we present CLUE, a bioinformatic and wet-lab pipeline for the multiplexed generation of pooled sgRNA libraries. CLUE starts from lists of genes or pasted sequences provided by the user and designs a single synthetic oligonucleotide pool containing various libraries. At the core of the approach, a barcoding

SUBMITTER: Becker M 

PROVIDER: S-EPMC7367185 | biostudies-literature | 2020 Jul

REPOSITORIES: biostudies-literature