RNA-programmed genome editing in human cells
Open accessAbstract
Type II CRISPR immune systems in bacteria use a dual RNA-guided DNA endonuclease, Cas9, to cleave foreign DNA at specific sites. We show here that Cas9 assembles with hybrid guide RNAs in human cells and can induce the formation of double-strand DNA breaks (DSBs) at a site complementary to the guide RNA sequence in genomic DNA. This cleavage activity requires both Cas9 and the complementary binding of the guide RNA. Experiments using extracts from transfected cells show that RNA expression and/or assembly into Cas9 is the limiting factor for Cas9-mediated DNA cleavage. In addition, we find that extension of the RNA sequence at the 3' end enhances DNA targeting activity in vivo. These results show that RNA-programmed genome editing is a facile strategy for introducing site-specific genetic changes in human cells.DOI:http://dx.doi.org/10.7554/eLife.00471.001.
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The CRISPR/Cas system has been successfully applied to human cells and several model organisms for genome editing.
“Recently, the CRISPR/Cas system has been successfully applied to human cells19,20,21,22 and several model organisms for genome editing.23,24,25,26”
From The CRISPR/Cas9 system facilitates clearance of the intrahepatic HBV templates in vivo (2014) · cites this paper as [22]
CRISPR/Cas9 is a naturally occurring bacterial immune system that protects bacteria from phage infection and has been harnessed for genome-engineering applications.
“Clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 is a naturally occurring bacterial immune system that uses a novel nuclease system to protect bacteria from phage infection (6-9), and it has recently been harnessed for a variety of genome-engineering applications (10)(11)(12)(13)(14)(15)(16)(17).”
From RNA-guided endonuclease provides a therapeutic strategy to cure latent herpesviridae infection (2014) · cites this paper as [12]
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- Authors
- Jinek, M., East, A., Cheng, A., Lin, S., Ma, E., & Doudna, J.
- Affiliations
- Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States
- University of California, Berkeley (US)
- Howard Hughes Medical Institute (US)
- Corresponding author
- Doudna, Jennifer
- Venue
- eLife
- Publisher
- eLife Sciences Publications, Ltd
- Published
- 2013-01-29
- Volume
- 2
- Pages
- e00471-e00471
- ISSN
- 2050-084X (ISSN-L)
- PMC
- PMC3557905
- OpenAlex
- W2153344788
- Cited by
- 2,098
- References
- 14
- Citations by year
- 2026: 36 · 2025: 83 · 2024: 104 · 2023: 123 · 2022: 174 · 2021: 159 · 2020: 191 · 2019: 181 · 2018: 196 · 2017: 203 · 2016: 226 · 2015: 174 · 2014: 153 · 2013: 95
- FWCI
- 96.82
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- eLife (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Zurich Open Repository and Archive (University of Zurich) (acceptedVersion, other-oa)
- OSTI OAI (U.S. Department of Energy Office of Scientific and Technical Information) (submittedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- eScholarship (California Digital Library) (submittedVersion)
- eScholarship (California Digital Library) (submittedVersion, cc-by)
- Zurich Open Repository and Archive (University of Zurich) (submittedVersion, other-oa)
- Zurich Open Repository and Archive (University of Zurich)
- Funders
- Howard Hughes Medical Institute
- National Institutes of Health
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- CRISPR/Cas9, Cas9, DNA double-strand breaks, site-specific DNA cleavage, DNA targeting, human cells, genome editing
- MeSH terms
- Humans, RNA, RNA Editing, Clustered Regularly Interspaced Short Palindromic Repeats
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