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Related Concept Videos

Prokaryotic Transcriptional Activators and Repressors01:58

Prokaryotic Transcriptional Activators and Repressors

The organization of prokaryotic genes in their genome is notably different from that of eukaryotes. Prokaryotic genes are organized, such that the genes for proteins involved in the same biochemical process or function are located together in groups. This group of genes, along with their regulatory elements, are collectively known as an operon. The functional genes in an operon are transcribed together to give a single strand of mRNA known as polycistronic mRNA.
Transcription of prokaryotic...
Prokaryotic Transcriptional Activators and Repressors01:58

Prokaryotic Transcriptional Activators and Repressors

The organization of prokaryotic genes in their genome is notably different from that of eukaryotes. Prokaryotic genes are organized, such that the genes for proteins involved in the same biochemical process or function are located together in groups. This group of genes, along with their regulatory elements, are collectively known as an operon. The functional genes in an operon are transcribed together to give a single strand of mRNA known as polycistronic mRNA.
Transcription of prokaryotic...
Repressible Operon: trp Operon01:21

Repressible Operon: trp Operon

The trp operon in Escherichia coli exemplifies a repressible operon. It regulates the synthesis of tryptophan through repressor-mediated transcriptional control and attenuation. This dual regulatory mechanism ensures tryptophan biosynthesis occurs only when needed, conserving cellular resources.Structure of the trp OperonThe trp operon consists of five structural genes (trpE, trpD, trpC, trpB, and trpA) that encode enzymes for tryptophan biosynthesis. These genes are transcribed as a single...
Types of RNA01:23

Types of RNA

Overview
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...
Constitutive and Regulated Gene Expression01:27

Constitutive and Regulated Gene Expression

Gene expression in prokaryotes is governed by constitutive and regulated systems, allowing cells to balance the production of essential proteins with adaptive responses to environmental changes.Constitutive Gene ExpressionConstitutive, or housekeeping, genes are continuously expressed as they encode proteins vital for fundamental cellular processes. These include enzymes for glycolysis, ribosomal components for protein synthesis, and proteins involved in DNA replication. Their constant...
Operons02:09

Operons

Prokaryotes can control gene expression through operons—DNA sequences consisting of regulatory elements and clustered, functionally related protein-coding genes. Operons use a single promoter sequence to initiate transcription of a gene cluster (i.e., a group of structural genes) into a single mRNA molecule. The terminator sequence ends transcription. An operator sequence, located between the promoter and structural genes, prohibits the operon’s transcriptional activity if bound by a repressor...

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Phage-mediated Delivery of Targeted sRNA Constructs to Knock Down Gene Expression in E. coli
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Artificial repressors for controlling gene expression in bacteria.

Mark C Politz1, Matthew F Copeland, Brian F Pfleger

  • 1University of Wisconsin-Madison, Department of Chemical and Biological Engineering, 3629 Engineering Hall, 1415 Engineering Dr Madison, WI, 53706, USA.

Chemical Communications (Cambridge, England)
|December 12, 2012
PubMed
Summary

Engineered transcription activator-like effector (TALE) proteins offer superior gene control in synthetic biology. These TALE proteins effectively block gene transcription, outperforming traditional methods like LacI for precise gene expression regulation.

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Area of Science:

  • Synthetic biology
  • Molecular biology
  • Gene expression regulation

Background:

  • Transcriptional repression is a key tool for controlling gene expression.
  • Existing methods like LacI have limitations in efficiency and specificity.
  • Need for novel repressors in synthetic biology applications.

Purpose of the Study:

  • To engineer and evaluate a novel DNA-binding protein for transcriptional repression.
  • To compare the efficacy of the engineered repressor against LacI.
  • To demonstrate the application of the engineered repressor in an operon context.

Main Methods:

  • Design and construction of a transcription activator-like effector (TALE)-based DNA-binding protein.
  • Assay of promoter blocking activity in vitro and in vivo.
  • Evaluation of gene repression in a downstream gene within an operon.

Main Results:

  • The engineered TALE-based protein demonstrated superior repression compared to LacI.
  • Effective blocking of transcription initiation from the target promoter was observed.
  • Successful repression of a downstream gene's expression within an operon was achieved.

Conclusions:

  • Engineered TALE proteins represent a powerful new tool for transcriptional repression in synthetic biology.
  • TALE-based repressors offer enhanced performance over existing systems like LacI.
  • This technology enables more precise control over gene expression in complex genetic circuits.