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 Experiment Videos

Improved ecdysone receptor-based inducible gene regulation system.

Subba R Palli1, Mariana Z Kapitskaya, Mohan B Kumar

  • 1Department of Entomology, College of Agriculture, University of Kentucky, KY 40546, USA. RPALLI@UKY.EDU

European Journal of Biochemistry
|March 13, 2003
PubMed
Summary

Researchers developed a novel ecdysone receptor (EcR)-based inducible gene system for precise control. This system demonstrates rapid and robust gene activation and deactivation in mammalian cells, offering significant potential for gene regulation applications.

Related Concept Videos

You might also read

Related Articles

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

Sort by
Same author

County-level surveillance for the American dog tick (Dermacentor variabilis) and Rickettsia species in Kentucky.

Scientific reports·2026
Same author

Histone acetylation homeodynamics navigates cell survival and apoptosis.

Nature communications·2025
Same author

20E-induced <i>Kr-h1</i> expression facilitates developmental transitions depending on chromosome accessibility of <i>BR-C</i> enhancers.

Proceedings of the National Academy of Sciences of the United States of America·2025
Same author

Juvenile hormone membrane signaling phosphorylates USP and thus potentiates 20-hydroxyecdysone action in Drosophila.

Science bulletin·2022
Same author

County-level surveillance for the lone star tick, Amblyomma americanum, and its associated pathogen, Ehrlichia chaffeensis, in Kentucky.

Ticks and tick-borne diseases·2022
Same author

Mapping distributions of the Lyme disease vector, Ixodes scapularis, and spirochete, Borrelia burgdorferi, in Kentucky using passive and active surveillance.

Ticks and tick-borne diseases·2021

Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Biotechnology

Background:

  • Ecdysone receptor (EcR) systems are valuable tools for inducible gene regulation.
  • Developing highly efficient and responsive EcR-based systems is crucial for various biological applications.
  • Existing systems may have limitations in induction levels, response times, or background activity.

Purpose of the Study:

  • To engineer a novel ecdysone receptor (EcR)-based inducible gene regulation system.
  • To optimize the system for high induction ratios and rapid response kinetics in mammalian cells.
  • To evaluate the performance of different construct combinations for maximal efficacy.

Main Methods:

  • Constructs were created by fusing domains from Choristoneura fumiferana EcR (CfEcR), C. fumiferana ultraspiracle (CfUSP), and Mus musculus retinoid X receptor (MmRXR) to GAL4 DNA binding domain (DBD) or VP16 activation domain (AD).

Related Experiment Videos

  • These constructs were tested in mammalian cells for their ability to transactivate a luciferase reporter gene under the control of GAL4 response elements and a synthetic TATAA promoter.
  • The most effective combination (GAL4 DBD-CfEcR (DEF) and VP16 AD-MmRXR (EF)) was identified through comparative analysis.

    • Main Results:

    • The optimal construct combination exhibited minimal background reporter gene activity without ligand and significantly high activity in its presence.
    • Ligand-induced gene expression showed a rapid 16-fold increase within 3 hours, reaching 8942-fold by 48 hours.
    • Gene expression rapidly decreased upon ligand withdrawal, with a 50% reduction at 12 hours and 80% at 24 hours.

    Conclusions:

    • A highly efficient EcR-based inducible gene regulation system was successfully developed.
    • The optimized system demonstrates fast induction and turn-off kinetics with substantial fold-induction.
    • This system offers a powerful and precise tool for inducible gene expression in mammalian systems.