Sakuma Tetsushi, Yamamoto Takashi
Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, 1-3-1 Kagamiyama, Higashi-Hiroshima, Hiroshima, 739-8526, Japan.
Methods Mol Biol. 2017;1630:25-36. doi: 10.1007/978-1-4939-7128-2_2.
Transcription activator-like effector (TALE) nuclease (TALEN) is the second-generation genome editing tool consisting of TALE protein containing customizable DNA-binding repeats and nuclease domain of FokI enzyme. Each DNA-binding repeat recognizes one base of double-strand DNA, and functional TALEN can be created by a simple modular assembly of these repeats. To easily and efficiently assemble the highly repetitive DNA-binding repeat arrays, various construction systems such as Golden Gate assembly, serial ligation, and ligation-independent cloning have been reported. In this chapter, we summarize the current situation of these systems and publically available reagents and protocols, enabling optimal selection of best suited systems for every researcher who wants to utilize TALENs in various research fields.
转录激活样效应物（TALE）核酸酶（TALEN）是第二代基因组编辑工具，由包含可定制DNA结合重复序列的TALE蛋白和FokI酶的核酸酶结构域组成。每个DNA结合重复序列识别双链DNA的一个碱基，通过这些重复序列的简单模块化组装即可构建功能性TALEN。为了简便高效地组装高度重复的DNA结合重复序列阵列，已报道了多种构建系统，如金门组装、串联连接和不依赖连接的克隆。在本章中，我们总结了这些系统以及公开可用试剂和方案的现状，以便为每个希望在各个研究领域使用TALEN的研究人员提供最佳选择，使其能够最优地选择最适合的系统。
