CRISPR Library Screening in Cultured Cardiomyocytes.

TitleCRISPR Library Screening in Cultured Cardiomyocytes.
Publication TypeJournal Article
Year of Publication2022
AuthorsS DeLuca, and N Bursac
JournalMethods in Molecular Biology (Clifton, N.J.)
Volume2485
Start Page1
Pagination1 - 13
Date Published01/2022
Abstract

CRISPR-Cas9-based screening technologies enable precise, high-throughput genetic and epigenetic manipulation to study mechanisms of development and disease and identify new therapeutic targets. Here, we describe a general protocol for the generation of custom, pooled CRISPR sgRNA libraries for screening in cardiomyocyte cultures. This methodology can address a variety of lab-specific research questions in cardiomyocytes and other cell types, as the genes to be modified can be curated or whole genomes can be investigated. The use of lentiviral sgRNA delivery followed by high-throughput sequencing allows for rapid comparison and identification of candidate genes and epigenetic modifiers, which can be further validated individually or in sub-pooled libraries following screening.

DOI10.1007/978-1-0716-2261-2_1
Short TitleMethods in Molecular Biology (Clifton, N.J.)