Targeted gene knockout by direct delivery of zinc-finger nuclease proteins
Open accessAbstract
Zinc-finger nucleases (ZFNs) are versatile reagents that have redefined genome engineering. Realizing the full potential of this technology requires the development of safe and effective methods for delivering ZFNs into cells. We demonstrate the intrinsic cell-penetrating capabilities of the standard ZFN architecture and show that direct delivery of ZFNs as proteins leads to efficient endogenous gene disruption in various mammalian cell types with minimal off-target effects.
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Gaj et al. found that the ZFP has an intrinsic cell-penetrating property that allows entry of the protein into cells without need for a delivery vehicle.
“Another exciting development in this regard is the finding by Gaj et al. that the ZFP has an intrinsic cell-penetrating property that allows entry of the protein into cells without need for a delivery vehicle.”
From Newer Gene Editing Technologies toward HIV Gene Therapy (2013) · cites this paper as [60]
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References 20 · Cited by 306
Research works printed but not linked: 1
- [16]Pruett-Miller SM Reading DW Porter SN Porteus MH PLoS Genet 5 e1000376 2009 19214211 PMC2633050
- Authors
- Gaj, T., Guo, J., Kato, Y., Sirk, S. J., & Barbas, C. F.
- Affiliations
- Scripps Research Institute (US)
- Corresponding author
- Barbas, Carlos F
- Venue
- Nature Methods
- Published
- 2012-07-01
- Volume
- 9
- Issue
- 8
- Pages
- 805-807
- ISSN
- 1548-7091 (ISSN-L)
- 1548-7105
- PMC
- PMC3424280
- OpenAlex
- W2000625273
- Cited by
- 306
- References
- 20
- Citations by year
- 2026: 6 · 2025: 6 · 2024: 13 · 2023: 12 · 2022: 19 · 2021: 10 · 2020: 13 · 2019: 20 · 2018: 23 · 2017: 25 · 2016: 35 · 2015: 44 · 2014: 42 · 2013: 30 · 2012: 5
- FWCI
- 15.54
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Nature Methods (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institutes of Health (DP1OD006990)
- Japan Society for the Promotion of Science
- National Cancer Institute (DP1CA174426)
- National Institute of General Medical Sciences (GM065059, T32GM080209, R01GM065059)
- 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 › Genetics › Virus-based gene therapy research
- Keywords
- zinc finger nucleases, protein delivery, off-target effects, genome editing
- MeSH terms
- Amino Acid Sequence, Animals, Cell Membrane Permeability, Genetic Engineering, Cricetinae, Humans, Molecular Sequence Data, Peptides, Recombinant Fusion Proteins, Deoxyribonucleases, Type II Site-Specific, Zinc Fingers, CHO Cells, Receptors, CCR5, Catalytic Domain, Amino Acid Motifs, tat Gene Products, Human Immunodeficiency Virus, Gene Knockout Techniques
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