title: Targeted transcriptional repression using a chimeric TALE-SRDX repressor protein.
abstract: 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.
authors.affiliation: Center for Plant Stress Genomics and Technology, King Abdullah University of Science and Technology, Thuwal 23955-6900, Kingdom of Saudi Arabia. magdy.mahfouz@kaust.edu.sa
authors.forename: Magdy M
authors.initials: MM
authors.lastname: Mahfouz
authors.forename: Lixin
authors.initials: L
authors.lastname: Li
authors.forename: Marek
authors.initials: M
authors.lastname: Piatek
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Fang
authors.forename: Hicham
authors.initials: H
authors.lastname: Mansour
authors.forename: Dhinoth K
authors.initials: DK
authors.lastname: Bangarusamy
authors.forename: Jian-Kang
authors.initials: JK
authors.lastname: Zhu
mesh_terms: D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D018744:DNA, Plant / Q000235:genetics; D017343:Genes, Plant; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D018744:DNA, Plant; C520629:RD29a protein, Arabidopsis; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins