Cell
Volume 154, Issue 2, 18 July 2013, Pages 442-451
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CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes

https://doi.org/10.1016/j.cell.2013.06.044Get rights and content
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Highlights

  • CRISPRi enables robust gene repression and activation in human cells

  • CRISPRi knockdown is specific with minimal off-target effects in human cells

  • CRISPRi can effectively repress endogenous genes in human and yeast

  • dCas9 enables modular and programmable RNA-guided genome regulation in eukaryotes

Summary

The genetic interrogation and reprogramming of cells requires methods for robust and precise targeting of genes for expression or repression. The CRISPR-associated catalytically inactive dCas9 protein offers a general platform for RNA-guided DNA targeting. Here, we show that fusion of dCas9 to effector domains with distinct regulatory functions enables stable and efficient transcriptional repression or activation in human and yeast cells, with the site of delivery determined solely by a coexpressed short guide (sg)RNA. Coupling of dCas9 to a transcriptional repressor domain can robustly silence expression of multiple endogenous genes. RNA-seq analysis indicates that CRISPR interference (CRISPRi)-mediated transcriptional repression is highly specific. Our results establish that the CRISPR system can be used as a modular and flexible DNA-binding platform for the recruitment of proteins to a target DNA sequence, revealing the potential of CRISPRi as a general tool for the precise regulation of gene expression in eukaryotic cells.

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