Center for Plant Stress Genomics and Technology, King Abdullah University of Science and Technology, Thuwal 23955-6900, Kingdom of Saudi Arabia.
Plant Mol Biol. 2012 Feb;78(3):311-21. doi: 10.1007/s11103-011-9866-x. Epub 2011 Dec 14.
Transcriptional activator-like effectors (TALEs) are proteins secreted by Xanthomonas bacteria when they infect plants. TALEs contain a modular DNA binding domain that can be easily engineered to bind any sequence of interest, and have been used to provide user-selected DNA-binding modules to generate chimeric nucleases and transcriptional activators in mammalian cells and plants. Here we report the use of TALEs to generate chimeric sequence-specific transcriptional repressors. The dHax3 TALE was used as a scaffold to provide a DNA-binding module fused to the EAR-repression domain (SRDX) to generate a chimeric repressor that targets the RD29A promoter. The dHax3.SRDX protein efficiently repressed the transcription of the RD29A::LUC transgene and endogenous RD29A gene in Arabidopsis. Genome wide expression profiling showed that the chimeric repressor also inhibited the expression of several other genes that contain the designer TALE-target sequence in their promoters. Our data suggest that TALEs can be used to generate chimeric repressors to specifically repress the transcription of genes of interest in plants. This sequence-specific transcriptional repression by direct on promoter effector technology is a powerful tool for functional genomics studies and biotechnological applications.
转录激活样效应因子（TALEs）是黄单胞菌在感染植物时分泌的蛋白质。TALEs 包含一个模块化的 DNA 结合结构域，该结构域可以很容易地被工程化为结合任何感兴趣的序列，并已被用于为哺乳动物细胞和植物提供用户选择的 DNA 结合模块，以生成嵌合核酸酶和转录激活因子。在这里，我们报告了使用 TALEs 生成嵌合序列特异性转录抑制剂。以 dHax3 TALE 为支架，提供与 EAR 抑制结构域（SRDX）融合的 DNA 结合模块，生成靶向 RD29A 启动子的嵌合抑制剂。dHax3.SRDX 蛋白有效地抑制了拟南芥中 RD29A::LUC 转基因和内源性 RD29A 基因的转录。全基因组表达谱分析表明，该嵌合抑制剂还抑制了启动子中含有设计的 TALE 靶序列的其他几个基因的表达。我们的数据表明，TALEs 可用于生成嵌合抑制剂，以特异性抑制植物中感兴趣基因的转录。这种通过直接在启动子上的效应物技术进行的序列特异性转录抑制是功能基因组学研究和生物技术应用的有力工具。
