Continuous directed evolution of DNA-binding proteins to improve TALEN specificity.
Continuous directed evolution of DNA-binding proteins to improve TALEN specificity.
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DOI：
10.1038/nmeth.3515
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发表时间：
2015-10
期刊：
影响因子：
48
通讯作者：
Liu DR
中科院分区：
文献类型：
--
作者：
Hubbard BP;Badran AH;Zuris JA;Guilinger JP;Davis KM;Chen L;Tsai SQ;Sander JD;Joung JK;Liu DR
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剂。在这里，我们提出了DNA结合噬菌体辅助连续进化（DB-PACE）作为DNA结合活性和特异性的实验室进化的一般方法。我们使用该系统来产生具有广泛改进的DNA切割特异性的TALE核酸酶，从而将DB-PACE确立为用于提高基因组编辑剂的准确性的通用方法。
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 TALE nucleases with broadly improved DNA cleavage specificity, establishing DB-PACE as a versatile approach for improving the accuracy of genome-editing agents.