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Related Experiment Videos

Targeting expression with light using caged DNA.

W T Monroe1, M M McQuain, M S Chang

  • 1Department of Biomedical Engineering, Vanderbilt University, Nashville, Tennessee 37235, USA.

The Journal of Biological Chemistry
|July 20, 1999
PubMed
Summary
This summary is machine-generated.

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Researchers developed a method to control gene expression using light. Caged plasmids were inactivated until exposed to laser light, enabling targeted gene expression with precision.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Plasmid-based gene expression is crucial in molecular biology.
  • Controlling gene expression spatially and temporally remains a challenge.
  • Photosensitive compounds offer potential for precise biological control.

Purpose of the Study:

  • To develop a method for light-inducible control of plasmid gene expression.
  • To demonstrate site-specific and temporal control of gene expression.
  • To investigate the mechanism of photosensitive caging on DNA transcription.

Main Methods:

  • DNA plasmids encoding luciferase and green fluorescent protein (GFP) were caged with 1-(4, 5-dimethoxy-2-nitrophenyl)diazoethane (DMNPE).
  • Caged plasmids were transfected into rat skin sites via particle bombardment and into HeLa cells via liposomes.

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  • Gene expression was induced and monitored upon exposure to 355-nm laser light.

    • Main Results:

    • Transfected skin sites with caged plasmids showed no luciferase expression.
    • Exposure to 355-nm laser light induced luciferase expression in a dose-dependent manner.
    • DMNPE caging blocked transcription of GFP plasmids in vitro, which was reversible with light exposure.
    • Caging altered plasmid electrophoretic mobility and optical absorption characteristics.

    Conclusions:

    • Photosensitive caging of plasmids with DMNPE allows for inactivation and site-specific, light-induced gene expression.
    • This technique offers precise spatial and temporal control over genetic material expression.
    • The method impacts gene expression at the transcriptional level and modifies plasmid properties.