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Related Experiment Videos

Direct gene transfer into the mouse heart

K Li1, R E Welikson, K L Vikstrom

  • 1Department of Microbiology and Immunology, Albert Einstein College of Medicine, NY, USA.

Journal of Molecular and Cellular Cardiology
|May 1, 1997
PubMed
Summary

Researchers developed a straightforward method for direct plasmid DNA injection into mouse hearts, enabling reproducible gene transfer for cardiac studies. This technique allows for stable gene expression and is valuable for screening transgenes before creating transgenic mice.

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Area of Science:

  • Cardiovascular Biology
  • Molecular Biology
  • Gene Therapy

Background:

  • Direct plasmid DNA injection into the myocardium is established for cardiac gene expression studies in various species.
  • Gene transfer into the mouse heart using plasmid DNA has not been previously reported, despite the importance of mouse models in genetic research.

Purpose of the Study:

  • To establish and validate a simple, reproducible method for direct gene transfer into the mouse heart using plasmid DNA.
  • To assess the characteristics of gene transfer, including stability, dose-dependency, spatial distribution, and suitability for different gene constructs.

Main Methods:

  • Direct injection of plasmid DNA encoding firefly luciferase (driven by RSV promoter) into the mouse heart.
  • Quantitative assessment of luciferase gene expression to determine spatial and temporal characteristics.

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  • Evaluation of gene transfer for structural genes using an epitope-tagged myosin heavy chain construct (driven by rat alpha-myosin heavy chain promoter).
  • Immunohistochemistry was used to detect protein expression.

    • Main Results:

    • Stable luciferase gene expression was observed for up to 8 weeks post-injection.
    • Gene expression demonstrated a dose-dependent response to input DNA (0.3-3 micrograms).
    • Low inter-animal variability and restricted gene expression to the left ventricle near the injection site.
    • The method successfully detected expression of structural genes under cellular promoters.

    Conclusions:

    • Direct naked DNA injection into the mouse heart provides a highly reproducible method for gene expression using both viral and cellular promoters.
    • This technique offers an inexpensive and efficient approach for in vivo cardiac gene regulation studies.
    • It serves as a valuable tool for screening potential transgenes prior to the generation of transgenic mice.