Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Riboswitches01:56

Riboswitches

9.9K
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...
9.9K
Types of RNA01:23

Types of RNA

73.5K
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...
73.5K
Transcriptional Regulation: Riboswitches01:23

Transcriptional Regulation: Riboswitches

969
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...
969
Translational Regulation01:29

Translational Regulation

739
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,...
739
Gene Regulation in Microbial Communities: Quorum Sensing01:28

Gene Regulation in Microbial Communities: Quorum Sensing

800
Quorum sensing is a mechanism of bacterial communication that enables coordinated gene expression in response to changes in population density. This facilitates collective behaviors that enhance survival, resource acquisition, and ecological adaptation. This process relies on small signaling molecules called autoinducers that accumulate as bacterial populations grow. When a critical threshold concentration of autoinducers is reached, bacterial cells collectively modify gene expression,...
800
Prokaryotic Transcriptional Activators and Repressors01:58

Prokaryotic Transcriptional Activators and Repressors

25.9K
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...
25.9K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Tagging lipoproteins in mycobacteria via metabolic incorporation of modified fatty acids.

Methods in enzymology·2026
Same author

Measuring Membrane Fluidity in Live Mycobacteria Reveals Subcellular Lateral Variation And Pole-Selective Responses to Mycomembrane Perturbation.

ACS infectious diseases·2026
Same author

Targeting Mycolic Acid Biosynthesis with Cyclic Sulfamates: A New Strategy against <i>Mycobacterium tuberculosis</i>.

ACS infectious diseases·2025
Same author

Measuring membrane fluidity in live mycobacteria reveals subcellular lateral variation and pole-selective responses to mycomembrane perturbation.

bioRxiv : the preprint server for biology·2025
Same author

Metabolic Tagging Reveals Surface-Associated Lipoproteins in Mycobacteria.

ACS infectious diseases·2025
Same author

Identifying Inhibitor Targets in Mycobacteria by Activity-Based Probe Profiling.

Methods in molecular biology (Clifton, N.J.)·2025

Related Experiment Video

Updated: Mar 9, 2026

Measurement of Specific Mycobacterial Mistranslation Rates with Gain-of-function Reporter Systems
06:18

Measurement of Specific Mycobacterial Mistranslation Rates with Gain-of-function Reporter Systems

Published on: April 26, 2019

6.5K

Characterization of Engineered PreQ1 Riboswitches for Inducible Gene Regulation in Mycobacteria.

Erik R Van Vlack1, Shana Topp2, Jessica C Seeliger3

  • 1Department of Chemistry, Stony Brook University, Stony Brook, New York, USA.

Journal of Bacteriology
|January 11, 2017
PubMed
Summary

Engineered preQ1 riboswitches enable inducible gene regulation in mycobacteria, even with a novel second binding site. These riboswitches offer titratable control for gene knockdown applications in diverse organisms.

Keywords:
inducible gene regulationmycobacteriapreQ1riboswitch

More Related Videos

An Assay for Quantifying Protein-RNA Binding in Bacteria
07:02

An Assay for Quantifying Protein-RNA Binding in Bacteria

Published on: June 12, 2019

7.1K
Live Cell Fluorescence Microscopy to Observe Essential Processes During Microbial Cell Growth
07:28

Live Cell Fluorescence Microscopy to Observe Essential Processes During Microbial Cell Growth

Published on: November 24, 2017

16.8K

Related Experiment Videos

Last Updated: Mar 9, 2026

Measurement of Specific Mycobacterial Mistranslation Rates with Gain-of-function Reporter Systems
06:18

Measurement of Specific Mycobacterial Mistranslation Rates with Gain-of-function Reporter Systems

Published on: April 26, 2019

6.5K
An Assay for Quantifying Protein-RNA Binding in Bacteria
07:02

An Assay for Quantifying Protein-RNA Binding in Bacteria

Published on: June 12, 2019

7.1K
Live Cell Fluorescence Microscopy to Observe Essential Processes During Microbial Cell Growth
07:28

Live Cell Fluorescence Microscopy to Observe Essential Processes During Microbial Cell Growth

Published on: November 24, 2017

16.8K

Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Synthetic Biology

Background:

  • Riboswitches are mRNA elements regulating gene expression in response to metabolites.
  • Mycobacteria lack endogenous preQ1 biosynthetic genes, suggesting potential for exogenous control.
  • Existing riboswitches offer tools for inducible gene expression across various organisms.

Purpose of the Study:

  • To investigate the behavior of naturally occurring and engineered preQ1 riboswitches in mycobacteria.
  • To apply preQ1 riboswitches for exogenous, inducible gene regulation in mycobacteria.
  • To engineer riboswitches for enhanced response ratios, repression, and gene knockdown applications.

Main Methods:

  • Assayed naturally occurring preQ1 riboswitches for gene repression in *Mycobacterium smegmatis*.
  • Employed structure-based design to engineer preQ1 riboswitches from *T. tencongensis*, *B. subtilis*, and *L. rhamnosus*.
  • Utilized reporter gene assays and dose-response modeling to characterize riboswitch performance.

Main Results:

  • Successfully demonstrated preQ1-dependent repression of a reporter gene in *M. smegmatis*.
  • Engineered riboswitches showed improved response ratios, though repression remained incomplete.
  • Discovered a novel, second independent preQ1 binding site influencing riboswitch dose response across all variants.

Conclusions:

  • Engineered preQ1 riboswitches provide a titratable and reversible system for inducible gene regulation in mycobacteria.
  • The identified second preQ1 binding site represents a previously undocumented aspect of riboswitch behavior.
  • Optimized riboswitches are suitable for inducible gene knockdown applications, expanding tools for mycobacterial research.