Akmammedov Arslan, Katsuyama Tomonori, Paro Renato
Department of Biosystems Science and Engineering, Eidgenössische Technische Hochschule Zürich, Mattenstrasse 26, 4058, Basel, Switzerland.
Department of Genetics, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo 7-3-1 Bunkyo-ku, 113-8654, Tokyo, Japan.
Methods Mol Biol. 2016;1480:269-81. doi: 10.1007/978-1-4939-6380-5_23.
Owing to their modular and highly specific DNA recognition mode, transcription activator-like effector nucleases (TALENs) have been rapidly adopted by the scientific community for the purpose of generating site-specific double-strand breaks (DSBs) on a DNA molecule. A pair of TALENs can be used to produce random insertions or deletions of various lengths via nonhomologous end-joining or together with a homologous donor DNA to induce precise sequence alterations by homologous recombination (HR). Here, we describe a method for TALEN assembly (easyT) and a strategy for genome engineering via HR.
由于其模块化和高度特异性的DNA识别模式，转录激活样效应物核酸酶（TALENs）已被科学界迅速采用，用于在DNA分子上产生位点特异性双链断裂（DSBs）。一对TALENs可用于通过非同源末端连接产生各种长度的随机插入或缺失，或与同源供体DNA一起通过同源重组（HR）诱导精确的序列改变。在这里，我们描述了一种TALEN组装方法（easyT）和一种通过HR进行基因组工程的策略。
