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Related Experiment Videos

Method for screening ecdysone-inducible stable cell lines.

K Wakita1, F McCormick, O Tetsu

  • 1University of California, San Francisco, San Francisco, CA, USA.

Biotechniques
|August 23, 2001
PubMed
Summary

Stable mammalian cell lines expressing high levels of ecdysone receptor proteins were generated using a luciferase reporter system. This method facilitates rapid screening and establishes reliable tools for gene function studies.

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Area of Science:

  • Mammalian cell biology
  • Molecular genetics
  • Gene expression regulation

Background:

  • Ecdysone-inducible systems are valuable for studying gene function in mammalian cells.
  • Establishing stable cell lines for these systems is challenging due to the need for multiple promoter-driven components.

Purpose of the Study:

  • To develop a rapid screening method for generating stable mammalian cell lines expressing ecdysone and retinoid X receptors.
  • To create reliable cell lines for conditional gene expression studies.

Main Methods:

  • Utilized a luciferase reporter gene system for efficient screening.
  • Generated two human stable colon cancer cell lines: SW480/VgRXR and HCT116/VgRXR.

Main Results:

Related Experiment Videos

  • Successfully established stable cell lines with sustained high-level expression of ecdysone receptor proteins for over six months.
  • Demonstrated tight, rapid, and persistent gene induction in SW480/VgRXR cells using 2 microM Ponasterone A.

    • Conclusions:

    • The luciferase reporter system offers an advantageous approach for rapidly generating stable cell lines with high ecdysone receptor expression.
    • These established cell lines provide robust tools for conditional gene function analysis in mammalian systems.