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

By Basil P. Hubbard, A. Badran, John A. Zuris, J. Guilinger, K. Davis, Chen Liwei, S. Tsai, Jeffry D. Sander, J. Keith Joung, David R. Liu. 2015

117 citations recorded by the source corpus.

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.

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Citations in the corpus

• Evolved Cas9 variants with broad PAM compatibility and high DNA specificity (2018, positive): “To link Cas9 DNA recognition to phage propagation during PACE, we developed a bacterial one-hybrid selection 20,23,24 in which the SP encodes a catalytically dead SpCas9 (dCas9) fused to the ω subunit of bacterial RNA polymerase.”