CRISPRi (CRISPR Interference)

Overview

CRISPRi uses a catalytically dead Cas9 (dCas9) fused to a transcriptional repressor domain (e.g., KRAB) guided by single-guide RNAs (sgRNAs) to silence target genomic loci without cutting DNA. Unlike canonical CRISPR-Cas9 editing, CRISPRi is reversible and does not introduce permanent sequence alterations. It is commonly applied to silence enhancers or promoters in a locus-specific manner, establishing causal links between non-coding regulatory elements and target gene expression.

Used by

• dCas9 + sgRNAs targeting the rs4519489 enhancer region used in 22Rv1 and PC3 prostate cancer cells; CRISPRi silencing of the rs4519489 locus significantly reduced NOL10 mRNA (P=9.79×10⁻⁵ and P=4.79×10⁻⁴ respectively), establishing a direct causal link between the enhancer and NOL10 expression PMID:28927585

Notes

• CRISPRi is locus-specific and reversible, making it well-suited to characterize the regulatory activity of non-coding elements without permanently altering the sequence.
• Complementary to CRISPR base editing, which permanently changes the allele.

Sources

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