CRISPR RNA–guided activation of endogenous human genes
Open accessAbstract
Short guide RNAs (gRNAs) can direct catalytically inactive CRISPR-associated 9 nuclease (dCas9) to repress endogenous genes in bacteria and human cells. Here we show that single or multiple gRNAs can direct dCas9 fused to a VP64 transcriptional activation domain to increase expression of endogenous human genes. This proof-of-principle work shows that clustered regularly interspaced short palindromic repeat (CRISPR)-Cas systems can target heterologous effector domains to endogenous sites in human cells.
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CRISPRi and CRISPRa technologies utilize catalytically inactive Cas9 (dCas9) fused to either a transcriptional repressor (e.g., KRAB) or a transcriptional activator (e.g., VPR).
“For example, CRISPRi (interference)23–25 and CRISPRa (activation)26–30 technologies utilize catalytically inactive Cas9 (dCas9) fused to either a transcriptional repressor (e.g., KRAB, Krüppel associated box) or a transcriptional activator (e.g., VPR, VP64-p65-Rta).”
From Large-scale CRISPR screening in primary human 3D gastric organoids enables comprehensive dissection of gene-drug interactions (2025) · cites this paper as [26]
The paper’s Abstract and Online Methods (page unavailable) describe catalytically inactive dCas9 fused to the VP64 activation domain to increase expression of endogenous human genes. The Abstract also mentions a dCas9-KRAB repressor fusion reported by another study. It does not describe a VPR (VP64-p65-Rta) activator, so the specific activator example is not supported by this source. (LLM, conf=0.86)
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- Authors
- Maeder, M. L., Linder, S. J., Cascio, V. M., Fu, Y., Ho, Q. H., & Joung, J. K.
- Affiliations
- Harvard University (US)
- Massachusetts General Hospital (US)
- Corresponding author
- Joung, J Keith
- Venue
- Nature Methods
- Published
- 2013-07-25
- Volume
- 10
- Issue
- 10
- Pages
- 977-979
- ISSN
- 1548-7091 (ISSN-L)
- 1548-7105
- PMC
- PMC3794058
- OpenAlex
- W2075230442
- Cited by
- 1,249
- References
- 7
- Citations by year
- 2026: 50 · 2025: 75 · 2024: 93 · 2023: 82 · 2022: 102 · 2021: 93 · 2020: 87 · 2019: 113 · 2018: 119 · 2017: 109 · 2016: 131 · 2015: 109 · 2014: 72 · 2013: 12
- FWCI
- 48.41
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Methods (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Digital Access to Scholarship at Harvard (DASH) (Harvard University) (submittedVersion, cc-by)
- Funders
- National Human Genome Research Institute (P50HG005550)
- National Institute of General Medical Sciences (DP1GM105378)
- National Institute of Neurological Disorders and Stroke (R01NS073124)
- National Center for Chronic Disease Prevention and Health Promotion (DP1 GM105378)
- 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/Cas9, dCas9, transcriptional activation, synthetic transcription factors, human cells
- MeSH terms
- Bacterial Proteins, Humans, Recombinant Fusion Proteins, Ribonucleases, Streptococcus pyogenes, Transcriptional Activation, RNA Editing, Vascular Endothelial Growth Factor A, HEK293 Cells, RNA, Small Untranslated, Clustered Regularly Interspaced Short Palindromic Repeats
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