Abstract
Nucleases containing programmable DNA-binding domains can alter the genomes of model organisms and have the potential to become human therapeutics. Here we present DNA-binding phage-assisted continuous evolution (DB-PACE) as a general approach for the laboratory evolution of DNA-binding activity and specificity. We used this system to generate transcription activator-like effectors nucleases (TALENs) with broadly improved DNA cleavage specificity, establishing DB-PACE as a versatile approach for improving the accuracy of genome-editing agents.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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DNA-Binding Proteins / chemistry
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism*
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Deoxyribonucleases / metabolism*
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Directed Molecular Evolution / methods*
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Gene Targeting / methods
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High-Throughput Screening Assays / methods
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Humans
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Polycomb Repressive Complex 1 / genetics
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Polycomb Repressive Complex 1 / metabolism
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Protein Engineering / methods
Substances
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CBX8 protein, human
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DNA-Binding Proteins
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Polycomb Repressive Complex 1
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Deoxyribonucleases