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Targeted transcriptional repression using a chimeric TALE-SRDX repressor protein

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 genera...

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Autores principales: Mahfouz, Magdy M., Li, Lixin, Piatek, Marek, Fang, Xiaoyun, Mansour, Hicham, Bangarusamy, Dhinoth K., Zhu, Jian-Kang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Netherlands 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3259320/
https://www.ncbi.nlm.nih.gov/pubmed/22167390
http://dx.doi.org/10.1007/s11103-011-9866-x
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author Mahfouz, Magdy M.
Li, Lixin
Piatek, Marek
Fang, Xiaoyun
Mansour, Hicham
Bangarusamy, Dhinoth K.
Zhu, Jian-Kang
author_facet Mahfouz, Magdy M.
Li, Lixin
Piatek, Marek
Fang, Xiaoyun
Mansour, Hicham
Bangarusamy, Dhinoth K.
Zhu, Jian-Kang
author_sort Mahfouz, Magdy M.
collection PubMed
description 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11103-011-9866-x) contains supplementary material, which is available to authorized users.
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spelling pubmed-32593202012-01-31 Targeted transcriptional repression using a chimeric TALE-SRDX repressor protein Mahfouz, Magdy M. Li, Lixin Piatek, Marek Fang, Xiaoyun Mansour, Hicham Bangarusamy, Dhinoth K. Zhu, Jian-Kang Plant Mol Biol Article 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11103-011-9866-x) contains supplementary material, which is available to authorized users. Springer Netherlands 2011-12-14 2012 /pmc/articles/PMC3259320/ /pubmed/22167390 http://dx.doi.org/10.1007/s11103-011-9866-x Text en © The Author(s) 2011 https://creativecommons.org/licenses/by-nc/4.0/ This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.
spellingShingle Article
Mahfouz, Magdy M.
Li, Lixin
Piatek, Marek
Fang, Xiaoyun
Mansour, Hicham
Bangarusamy, Dhinoth K.
Zhu, Jian-Kang
Targeted transcriptional repression using a chimeric TALE-SRDX repressor protein
title Targeted transcriptional repression using a chimeric TALE-SRDX repressor protein
title_full Targeted transcriptional repression using a chimeric TALE-SRDX repressor protein
title_fullStr Targeted transcriptional repression using a chimeric TALE-SRDX repressor protein
title_full_unstemmed Targeted transcriptional repression using a chimeric TALE-SRDX repressor protein
title_short Targeted transcriptional repression using a chimeric TALE-SRDX repressor protein
title_sort targeted transcriptional repression using a chimeric tale-srdx repressor protein
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3259320/
https://www.ncbi.nlm.nih.gov/pubmed/22167390
http://dx.doi.org/10.1007/s11103-011-9866-x
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