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Related Concept Videos

Co-activators and Co-repressors02:04

Co-activators and Co-repressors

Gene transcription is regulated by the synergistic action of several proteins that form a complex at a gene regulatory site. This is observed in eukaryotes, where the regulation of gene expression is a complex process. Regulatory proteins in eukaryotes can broadly be classified into two types – regulators that bind directly to specific DNA sequences and co-regulators that associate with regulatory proteins but cannot directly bind to the DNA. These co-regulators are further divided into...
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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Eukaryotic Transcription Inhibitors

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Proteins that regulate transcription can do so either via direct contact with RNA Polymerase or through indirect interactions facilitated by adaptors, mediators, histone-modifying proteins, and nucleosome remodelers. Direct interactions to activate transcription is seen in bacteria as well as in some eukaryotic genes. In these cases, upstream activation sequences are adjacent to the promoters, and the activator proteins interact directly with the transcriptional machinery. For example, in...
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In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing
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Targeted transcriptional repression using a chimeric TALE-SRDX repressor protein.

Magdy M Mahfouz1, Lixin Li, Marek Piatek

  • 1Center 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

Plant Molecular Biology
|December 15, 2011
PubMed
Summary

Transcriptional activator-like effectors (TALEs) can be engineered to create sequence-specific repressors. This technology allows precise control over gene transcription in plants for research and biotechnology.

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Area of Science:

  • Plant molecular biology
  • Synthetic biology
  • Bacterial effector proteins

Background:

  • Transcriptional activator-like effectors (TALEs) are bacterial proteins with customizable DNA-binding domains.
  • TALEs have been previously engineered for gene activation and nuclease applications in various organisms.
  • Targeted gene repression in plants using engineered DNA-binding proteins is a valuable tool.

Purpose of the Study:

  • To engineer and characterize TALE-based chimeric repressors for sequence-specific gene silencing in plants.
  • To demonstrate the efficacy of TALE-SRDX repressors in targeting specific promoters and endogenous genes.
  • To explore the potential of TALE technology for broad gene regulation applications in plants.

Main Methods:

  • Engineering a chimeric repressor by fusing a TALE DNA-binding module (dHax3) with the EAR repression domain (SRDX).
  • Testing the dHax3.SRDX repressor's activity on a RD29A promoter-driven reporter gene (LUC) and endogenous RD29A in Arabidopsis.
  • Performing genome-wide expression profiling to identify additional genes regulated by the chimeric repressor.

Main Results:

  • The engineered dHax3.SRDX protein effectively repressed transcription from the RD29A promoter in Arabidopsis.
  • Both RD29A::LUC transgene and endogenous RD29A gene expression were significantly inhibited by the dHax3.SRDX repressor.
  • Genome-wide analysis revealed that the chimeric repressor also targeted other genes containing the designed TALE-binding sequence in their promoters.

Conclusions:

  • TALEs can be successfully engineered into sequence-specific transcriptional repressors for use in plants.
  • This TALE-based repression system offers precise control over gene expression, enabling targeted functional genomics studies.
  • The developed technology holds promise for diverse biotechnological applications requiring specific gene silencing in plants.