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Related Experiment Videos

In vivo gene delivery and expression by bacteriophage lambda vectors.

H A Lankes1, C N Zanghi, K Santos

  • 1Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, NY 14642, USA.

Journal of Applied Microbiology
|April 24, 2007
PubMed
Summary
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Bacteriophage lambda can transfer genes into mammalian cells in vivo, with expression peaking within 24 hours. Surface modifications improve phage uptake and gene transfer efficiency in mammals.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Virology

Background:

  • Bacteriophages offer cost-effective, safe, and stable vectors for gene transfer and vaccine delivery.
  • Phage-mediated gene transfer in mammalian systems is not well understood.
  • Investigating phage gene transfer in vivo is crucial for developing novel therapeutic strategies.

Purpose of the Study:

  • To investigate the efficacy and mechanisms of phage-mediated gene transfer in vivo in mammalian hosts.
  • To assess the potential of bacteriophage lambda as a gene transfer vector in mice.
  • To explore strategies for enhancing phage-mediated gene transfer efficiency.

Main Methods:

  • Mice were inoculated with recombinant lambda phage carrying a firefly luciferase (luc) expression cassette.

Related Experiment Videos

  • In vivo gene expression was measured by luciferase activity over time.
  • Phage uptake was enhanced by displaying an integrin-binding peptide on the phage surface.
  • Phagocytic cells were depleted using clodronate liposomes to assess their role in gene transfer.

    • Main Results:

    • Efficient, dose-dependent in vivo luciferase expression was observed, peaking at 24 hours post-delivery.
    • Surface modification with an integrin-binding peptide increased phage internalization in vitro and gene transfer in vivo.
    • Depletion of phagocytic cells had a minimal impact on the efficiency of phage-mediated gene transfer.
    • Unmodified lambda phage particles demonstrated the ability to transduce mammalian cells in vivo.

    Conclusions:

    • Lambda phage can effectively transduce mammalian cells in vivo, suggesting potential as a gene delivery vector.
    • Gene transfer may occur via non-phagocytic uptake mechanisms.
    • Surface modifications enhancing phage uptake significantly improve in vivo gene transfer efficiency.
    • These findings provide insights into phage-mediated gene transfer mechanisms and suggest avenues for enhancing therapeutic applications.