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Related Concept Videos

Types of RNA01:23

Types of RNA

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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.
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Riboswitches01:56

Riboswitches

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Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
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Transcriptional Regulation: Riboswitches01:23

Transcriptional Regulation: Riboswitches

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Riboswitches are RNA elements that regulate gene expression by altering their secondary structures in response to specific effector molecules. These elements, located in the leader regions of certain mRNAs, act as transcriptional regulators by toggling between alternative conformations to control downstream gene expression. Riboswitch-mediated regulation is a precise mechanism for modulating biosynthetic pathways, as exemplified by the riboflavin biosynthesis pathway in Bacillus...
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Translational Regulation01:29

Translational Regulation

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Translational regulation in prokaryotes ensures efficient protein synthesis by controlling ribosome access to mRNA. This regulation is mediated by secondary RNA structures, including translational riboswitches, RNA thermometers, and small RNAs (sRNAs), which respond to intracellular and environmental signals to modulate gene expression.Translational RiboswitchesRiboswitches in the leader region of mRNAs can regulate translation by altering the accessibility of the Shine-Dalgarno (SD) sequence,...
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Repressible Operon: trp Operon01:21

Repressible Operon: trp Operon

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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...
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Reporter Genes02:11

Reporter Genes

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Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
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Related Experiment Video

Updated: Jul 11, 2025

Applying an Inducible Expression System to Study Interference of Bacterial Virulence Factors with Intracellular Signaling
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Applying an Inducible Expression System to Study Interference of Bacterial Virulence Factors with Intracellular Signaling

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Engineering U1-Based Tetracycline-Inducible Riboswitches to Control Gene Expression in Mammals.

Eric Rovira1, Beatriz Moreno2, Nerea Razquin1

  • 1Department of Gene Therapy and Regulation of Gene Expression, Center for Applied Medical Research (CIMA), University of Navarra (UNAV), Pamplona 31008, Spain.

ACS Nano
|November 16, 2023
PubMed
Summary

Synthetic riboswitches offer precise gene control. Researchers developed tetracycline-inducible U1snRNP interference (U1i) riboswitches, enhanced by Systematic Evolution of Riboswitches by Exponential Enrichment (SEREX), achieving significant gene expression control for potential gene therapy.

Keywords:
SELEXU1 snRNPU1iaptamergene expression regulationriboswitchtetracycline

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Last Updated: Jul 11, 2025

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Area of Science:

  • Synthetic biology
  • Molecular biology
  • Gene regulation

Background:

  • Synthetic riboswitches are valuable tools for fine-tuning gene expression due to their small size and lack of immunogenicity.
  • Existing systems often require exogenous trans-acting factors for regulation.
  • U1snRNP interference (U1i) offers a novel gene inhibition mechanism.

Purpose of the Study:

  • To engineer tetracycline (TC)-inducible riboswitches based on the U1i system.
  • To enhance riboswitch activity and U1 snRNP binding capacity using Systematic Evolution of Riboswitches by Exponential Enrichment (SEREX).
  • To evaluate the potential of these riboswitches for gene therapy applications.

Main Methods:

  • Engineering of TC-U1i riboswitches to modulate mRNA polyadenylation via U1 snRNP recruitment.
  • Application of SEREX for isolating high-activity riboswitches.
  • Multiplexing of riboswitches to amplify induction levels.
  • Testing dose-dependency, reversibility, and regulation of reporter/endogenous genes in cellular and animal models.

Main Results:

  • Engineered TC-U1i riboswitches achieved 3-to-4-fold gene expression induction.
  • SEREX technology isolated riboswitches with up to 8-fold induction.
  • Multiplexing riboswitches resulted in up to 37-fold induction.
  • U1i-based riboswitches demonstrated dose-dependent, reversible regulation in vitro and in vivo.

Conclusions:

  • U1i-based riboswitches provide a robust platform for inducible gene expression control.
  • SEREX is an effective technology for discovering novel riboswitches with enhanced functionality.
  • These riboswitches show significant promise for gene therapy applications due to their efficacy and controllability.