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

Details

• Alternative Title:
  Nat Methods
• Personal Author:
  Hubbard, Basil P. ; Badran, Ahmed H. ; Zuris, John A. ; Guilinger, John P. ; Davis, Kevin M. ; Chen, Liwei ; Tsai, Shengdar Q. ; Sander, Jeffry D. ; Joung, J. Keith ; Liu, David R.
• Description:
  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.
• Subjects:
  Article Deoxyribonucleases Directed Molecular Evolution DNA-Binding Proteins Gene Targeting High-Throughput Screening Assays Humans Polycomb Repressive Complex 1 Protein Engineering
• Source:
  Nat Methods. 12(10):939-942.
• Pubmed ID:
  26258293
• Pubmed Central ID:
  PMC4589463
• Document Type:
  Journal Article
• Funding:
  DP1 GM105378/DP/NCCDPHP CDC HHS/United States ; F32 GM105189/GM/NIGMS NIH HHS/United States ; R01 GM065400/GM/NIGMS NIH HHS/United States ; R01 GM065400/GM/NIGMS NIH HHS/United States ; R01 GM095501/GM/NIGMS NIH HHS/United States ; R01 GM095501/GM/NIGMS NIH HHS/United States ; T32 GM007598/GM/NIGMS NIH HHS/United States
• Volume:
  12
• Issue:
  10
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:e43c791f90b6ea50b8bd830f7cab0d4c5a9afe8ceac78e4d15d8a0b5e4948887
• Download URL:
  https://stacks.cdc.gov/view/cdc/36969/cdc_36969_DS1.pdf
• File Type:
  [PDF - 820.89 KB ]