Transcription activator-like effector (TALE) nucleases (TALENs) efficiently recognize and cleave DNA in a sequence-dependent manner. However, current TALE custom synthesis methods are either complicated or expensive. Here we report a simple and low-cost method for TALE construct assembly. This method utilizes the denaturation/reannealing nature of double-stranded DNA to create a unique single-stranded DNA overhang for proper ordering of TALE monomers in an engineered multimer. We successfully synthesized two TALEN pairs targeting the endogenous TET1 locus in human embryonic kidney cells and demonstrated their editing efficiency. Our method provides an alternative simple, low-cost method for effective TALEN assembly, which may improve the application of TALE-based technology.

中文翻译：

---

通过双引物组装简单高效的定制转录激活因子样效应子基因合成。

转录激活因子样效应子 (TALE) 核酸酶 (TALEN) 以依赖序列的方式有效识别和切割 DNA。然而，当前的 TALE 定制合成方法要么复杂要么昂贵。在这里，我们报告了一种用于 TALE 构造组装的简单且低成本的方法。该方法利用双链 DNA 的变性/重退火特性来创建独特的单链 DNA 突出端，以便在工程多聚体中正确排序 TALE 单体。我们成功合成了两个针对人胚胎肾细胞中内源性TET1基因座的TALEN 对，并证明了它们的编辑效率。我们的方法为有效的 TALEN 组装提供了一种替代的简单、低成本的方法，这可能会改进基于 TALE 的技术的应用。