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Related Experiment Videos

Selection for retroviral insertions into regulated genes

J A Gogos1, W Lowry, M Karayiorgou

  • 1Fred Hutchinson Center for Cancer Research, Seattle, Washington 98104, USA. gogosj@rockvax.rockefeller.edu

Journal of Virology
|February 1, 1997
PubMed
Summary
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This study introduces a novel gene trapping strategy to identify genes regulated during cellular processes. The method efficiently screens thousands of gene trap integrations to find genes activated by specific proteins like MyoD.

Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Gene traps are valuable tools for monitoring gene expression changes during cellular processes.
  • Identifying genes regulated by specific cellular signals remains a challenge.

Purpose of the Study:

  • To develop and demonstrate a strategy combining retroviral gene traps, advanced selection methods, and responsive cell lines.
  • To enrich for retroviral insertions into regulated genes, including those involved in differentiation and response to stimuli.
  • To identify genes directly or indirectly regulated by the MyoD protein.

Main Methods:

  • Utilized retroviral gene trap vectors for stable integration into the host genome.
  • Employed fluorescence-activated cell sorting and dominant drug selection for efficient enrichment of targeted insertions.

Related Experiment Videos

  • Established a MyoD-responsive fibroblast cell line to screen for genes activated by MyoD.
  • Rescued flanking sequences of gene trap integrations for host gene identification.

    • Main Results:

    • Successfully screened thousands of gene trap integrations in a single experiment.
    • Identified gene trap insertions representing direct or indirect targets of MyoD.
    • Observed distinct patterns of gene regulation during myogenic determination.

    Conclusions:

    • The presented gene trapping strategy is feasible and efficient for identifying regulated genes.
    • This approach facilitates the discovery of genes involved in cellular differentiation and response pathways.
    • The method provides a foundation for isolating host genes and advancing genome-wide sequencing projects.