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Nat Methods DOI:10.1038/nmeth.3515

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

Publication TypeJournal Article
Year of Publication2015
AuthorsHubbard, BP, Badran, AH, Zuris, JA, Guilinger, JP, Davis, KM, Chen, L, Tsai, SQ, Sander, JD, J Joung, K, Liu, DR
JournalNat Methods
Date Published2015 Oct
KeywordsDeoxyribonucleases, Directed Molecular Evolution, DNA-Binding Proteins, Gene Targeting, High-Throughput Screening Assays, Humans, Polycomb Repressive Complex 1, Protein Engineering

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.


Alternate JournalNat. Methods
PubMed ID26258293
PubMed Central IDPMC4589463
Grant ListDP1 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