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Nat Methods. 2015 Oct;12(10):939-42. doi: 10.1038/nmeth.3515. Epub 2015 Aug 10.

Continuous directed evolution of DNA-binding proteins to improve TALEN specificity.

Author information

1
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
2
Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, Massachusetts, USA.
3
Center for Cancer Research, Massachusetts General Hospital, Charlestown, Massachusetts, USA.
4
Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, Massachusetts, USA.
5
Department of Pathology, Harvard Medical School, Boston, Massachusetts, USA.
6
Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts, USA.

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.

PMID:
26258293
PMCID:
PMC4589463
DOI:
10.1038/nmeth.3515
[Indexed for MEDLINE]
Free PMC Article

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