Engineering novel CRISPRi repressors for highly efficient mammalian gene regulation.

ABSTRACT:

Background

CRISPR interference (CRISPRi), the repurposing of the RNA-guided endonuclease dCas9 as a programmable transcriptional repressor, allows highly specific repression (knockdown) of gene expression. CRISPRi platforms can often have incomplete knockdown, performance variability across cell lines and gene targets, and inconsistencies dependent on the guide RNA sequence employed.

Results

Here, we explore the combination of novel repressor domains with strong Krüppel-associated box (KRAB) repressors, screening > 100 bipartite and tripartite fusion proteins for their ability to reduce gene expression as CRISPRi effectors. We show that these novel repressor fusions have reduced dependence on guide RNA sequences, better slow cell growth when used to knock down expression of

SUBMITTER: Kristof A 

PROVIDER: S-EPMC12164210 | biostudies-literature | 2025 Jun

REPOSITORIES: biostudies-literature