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

Nat Methods
Authors
Keywords
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.

Year of Publication
2015
Journal
Nat Methods
Volume
12
Issue
10
Pages
939-42
Date Published
2015 Oct
ISSN
1548-7105
DOI
10.1038/nmeth.3515
PubMed ID
26258293
PubMed Central ID
PMC4589463
Links
Grant list
DP1 GM105378 / GM / NIGMS NIH HHS / United States
T32 GM007598 / GM / NIGMS NIH HHS / United States
R01 GM065400 / GM / NIGMS NIH HHS / United States
DP1 GM105378 / DP / NCCDPHP CDC HHS / United States
F32 GM105189 / GM / NIGMS NIH HHS / United States
R01 GM095501 / GM / NIGMS NIH HHS / United States