Abstract
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.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Motifs
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Amino Acid Sequence
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Animals
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CHO Cells
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Catalytic Domain
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Cell Membrane Permeability
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Cricetinae
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Deoxyribonucleases, Type II Site-Specific / chemistry
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Deoxyribonucleases, Type II Site-Specific / metabolism*
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Gene Knockout Techniques / methods*
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Genetic Engineering / methods*
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Humans
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Molecular Sequence Data
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Peptides / chemistry
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Receptors, CCR5 / genetics
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism*
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Zinc Fingers / genetics
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Zinc Fingers / physiology*
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tat Gene Products, Human Immunodeficiency Virus / chemistry
Substances
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Peptides
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Receptors, CCR5
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Recombinant Fusion Proteins
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tat Gene Products, Human Immunodeficiency Virus
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polyarginine
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endodeoxyribonuclease FokI
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Deoxyribonucleases, Type II Site-Specific