清脆的
亚基因组mRNA
计算生物学
表观遗传学
鉴定(生物学)
生物
Cas9
高通量筛选
基因组
基因
遗传学
植物
作者
Sophia DeLuca,Nenad Bursac
出处
期刊:Methods in molecular biology
日期:2022-01-01
卷期号:: 1-13
标识
DOI:10.1007/978-1-0716-2261-2_1
摘要
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.
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