Cited by 3,696CRISPR and Genetic EngineeringRNA Interference and Gene DeliveryRNA regulation and disease

The DOI matches its Crossref record; 46 of the 46 references it cites were found in a registry or an index; none of the 5 sources its claims cite is checked for support yet.

CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes

Open access

Abstract

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.

Overview

Integrity signals

Registry

Crossref · 10.1016/j.cell.2013.06.044

Cites

46 of 46 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Verification

Claims the paper makes

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Show all 36 claims

  • It may state two separate claims as one. 14
  • Probably not a substantial claim. 7
20 of 20
  • The paper relies on this without support.

    The fluorescent antibodies used to stain CD71 and CXCR4 are specific for those proteins.

    “We designed sgRNAs to target genes encoding cell surface transmembrane proteins, enabling us to use flow cytometry with directly conjugated fluorescent antibodies to quantify gene expression at the single-cell level.”
  • The paper relies on this without support.

    Only a small fraction of the human genome is transcribed or required for controlling transcription.

    “only a small fraction of the human genome is transcribed or required for controlling transcription”
  • The paper relies on this without support.

    CRISPRi acts at the DNA level.

    “because CRISPRi acts at the DNA level”
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Metadata

Authors
Gilbert, L. A., Larson, M. H., Morsut, L., Liu, Z., Brar, G. A., Torres, S. E., Stern-Ginossar, N., Brandman, O., Whitehead, E. H., Doudna, J. A., Lim, W. A., Weissman, J. S., & Qi, L. S.
Affiliations
  • QB3 (US)
  • Howard Hughes Medical Institute (US)
  • Peking University (CN)
Corresponding author
Weissman, Jonathan S.
Venue
Cell
Publisher
Elsevier BV
Published
2013-07
Volume
154
Issue
2
Pages
442-451
ISSN
0092-8674 (ISSN-L)
OpenAlex
W2101749257
Cited by
3,696
References
46
Citations by year
2026: 195 · 2025: 260 · 2024: 298 · 2023: 286 · 2022: 316 · 2021: 374 · 2020: 387 · 2019: 332 · 2018: 328 · 2017: 286 · 2016: 256 · 2015: 187 · 2014: 161 · 2013: 28
FWCI
94.43
Type
journal article
Language
en
Open access
Locations
  • Cell (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
  • eScholarship (California Digital Library) (submittedVersion)
Funders
  • National Science Foundation (U01 CA168370, P50 GM102706, 0540879)
  • Howard Hughes Medical Institute
  • National Institutes of Health (GM102706, GM081879)
  • University of California, San Francisco (GM081879)
  • National Cancer Institute (U01CA168370)
  • National Institute of General Medical Sciences (P50GM081879, P50GM102706, GM081879, F32GM100611, GM102706)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
Keywords
CRISPR interference, dCas9, transcriptional repression, transcriptional activation, gene silencing, CRISPR/Cas9
MeSH terms
Bacterial Proteins, HeLa Cells, Humans, Saccharomyces cerevisiae, Streptococcus pyogenes, Gene Targeting, HEK293 Cells, RNA, Small Untranslated

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