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
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, Massachusetts, USA.
Nat Methods. 2015 Oct;12(10):939-42. doi: 10.1038/nmeth.3515. Epub 2015 Aug 10.
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.
含有可编程DNA结合结构域的核酸酶可以改变模式生物的基因组，并有可能成为人类治疗药物。在此，我们提出了DNA结合噬菌体辅助连续进化（DB-PACE），作为一种用于DNA结合活性和特异性实验室进化的通用方法。我们使用该系统生成了具有广泛改善的DNA切割特异性的转录激活样效应物核酸酶（TALENs），确立了DB-PACE作为提高基因组编辑试剂准确性的通用方法。
