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Related Experiment Videos

Imaging transgene expression in live animals.

A Honigman1, E Zeira, P Ohana

  • 1Department of Virology, Faculty of Medicine, Hebrew University, Jerusalem, Israel.

Molecular Therapy : the Journal of the American Society of Gene Therapy
|September 8, 2001
PubMed
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A new cooled charged-coupled device (CCCD) camera enables continuous in vivo monitoring of gene therapy delivery systems in living animals. This method allows real-time assessment of therapeutic gene expression across various tissues without sacrificing the subjects.

Area of Science:

  • Biomedical Engineering
  • Molecular Biology
  • Gene Therapy

Background:

  • Assessing therapeutic gene expression in disease models is crucial for evaluating gene therapy efficacy.
  • Current methods often require animal sacrifice, limiting continuous monitoring.
  • Non-invasive, real-time assessment tools are needed for in vivo gene therapy studies.

Purpose of the Study:

  • To introduce and evaluate a novel light-detection cooled charged-coupled device (CCCD) camera system.
  • To demonstrate its capability for continuous in vivo monitoring of gene expression.
  • To assess the efficiency and specificity of various gene delivery systems in living animals.

Main Methods:

  • Application of a CCCD camera for real-time, in vivo monitoring of luciferase (luc) gene expression.

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  • Testing diverse gene therapy delivery systems: ex vivo cells, viral vectors, and naked DNA.
  • Evaluation across multiple tissues (bone, muscle, liver, etc.) in mice and rats using various delivery methods (electroporation, injection).

    • Main Results:

    • The CCCD camera provides rapid detection (approx. 2 minutes exposure) of luc activity compared to older methods (approx. 15 minutes).
    • Successfully monitored gene expression from various vectors and delivery methods in multiple organs.
    • Confirmed luciferin distribution is not a limitation for in vivo detection.

    Conclusions:

    • The CCCD photon detection system offers a simple, reproducible, and applicable method for continuous in vivo monitoring of gene delivery.
    • This technology facilitates real-time assessment of gene therapy effectiveness without animal sacrifice.
    • Enables detailed analysis of gene expression location, intensity, and duration in living subjects.