Optimizing sgRNA position markedly improves the efficiency of CRISPR/dCas9-mediated transcriptional repression.

ABSTRACT: CRISPR interference (CRISPRi) represents a newly developed tool for targeted gene repression. It has great application potential for studying gene function and mapping gene regulatory elements. However, the optimal parameters for efficient single guide RNA (sgRNA) design for CRISPRi are not fully defined. In this study, we systematically assessed how sgRNA position affects the efficiency of CRISPRi in human cells. We analyzed 155 sgRNAs targeting 41 genes and found that CRISPRi efficiency relies heavily on the precise recruitment of the effector complex to the target gene transcription start site (TSS). Importantly, we demonstrate that the FANTOM5/CAGE promoter atlas represents the most reliable source of TSS annotations for this purpose. We also show that the proximity to the FANTOM5/CAGE

SUBMITTER: Radzisheuskaya A 

PROVIDER: S-EPMC5062975 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature