Inducible modulation is often required for precise investigations and manipulations of dynamic biological processes. Transcription activator-like effectors (TALEs) provide a powerful tool for targeted gene editing and transcriptional programming. We designed a series of chemical inducible systems by coupling TALEs with a mutated human estrogen receptor (ER^T2), which renders them 4-hydroxyl-tamoxifen (4-OHT) inducible for access of the genome. Chemical inducible genome editing was achieved via fusing two tandem ER^T2 domains to customized transcription activator-like effector nuclease (TALEN), which we termed “Hybrid Inducible Technology” (HIT-TALEN). Those for transcription activation were vigorously optimized using multiple construct designs. Most efficient drug induction for endogenous gene activation was accomplished with minimal background activity using an optimized inducible TALE based SunTag system (HIT-TALE-SunTag). The HIT-SunTag system is rapid, tunable, selective to 4-OHT over an endogenous ligand, and reversible in drug induced transcriptional activation. Versatile systems developed in this study can be easily applied for editing and transcriptional programming of potentially any genomic loci in a tight and effective chemical inducible fashion.

中文翻译：

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用于基因组编辑和转录激活的多种化学诱导性Tal效应子

精确调节和动态生物学过程的操纵通常需要诱导调制。转录激活子样效应子（TALE）为目标基因编辑和转录编程提供了强大的工具。我们通过将TALE与突变的人类雌激素受体（ER ^T2）偶联，设计了一系列化学诱导系统，这使它们可以被4-羟基-他莫昔芬（4-OHT）诱导进入基因组。通过将两个串联的ER ^T2融合，实现了化学诱导的基因组编辑定制转录激活因子样效应物核酸酶（TALEN）的结构域，我们称之为“杂交诱导技术”（HIT-TALEN）。使用多种构建体设计大力优化了转录激活序列。使用优化的可诱导的基于TALE的SunTag系统（HIT-TALE-SunTag），以最小的背景活性完成了最有效的内源基因激活药物诱导。HIT-SunTag系统快速，可调，对内源性配体具有4-OHT选择性，并且在药物诱导的转录激活中可逆。在这项研究中开发的多功能系统可以很容易地以紧密和有效的化学诱导方式轻松地用于可能的任何基因组位点的编辑和转录程序设计。