Journal
CELL
Volume 154, Issue 2, Pages 442-451Publisher
CELL PRESS
DOI: 10.1016/j.cell.2013.06.044
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Funding
- UCSF Center for Systems and Synthetic Biology
- NIH [P50 GM102706, P50 GM081879, U01 CA168370]
- Howard Hughes Medical Institute
- NSF SynBERC [EEC-0540879]
- NIGMS IMSD
- Human Frontiers Scientific Program
- Ruth L. Kirschstein National Research Service Award
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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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