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Related Experiment Videos

Evolving phage vectors for cell targeted gene delivery.

David Larocca1, Michael A Burg, Kristen Jensen-Pergakes

  • 1Selective Genetics, Inc, San Diego, CA 92121, USA. laroccad@selectivegenetics.com

Current Pharmaceutical Biotechnology
|March 9, 2002
PubMed
Summary

Filamentous phage vectors were engineered for targeted gene delivery to mammalian cells. Modifications significantly improved transduction efficiency, showing promise for advanced gene therapy applications.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Gene Therapy

Background:

  • Filamentous phage are simple, economical biological entities with no intrinsic mammalian tropism.
  • Phage vectors can be genetically modified for targeted gene delivery, offering advantages over viral vectors.
  • Previous phage-mediated gene transfer showed low efficiency, necessitating optimization.

Purpose of the Study:

  • To adapt filamentous phage for targeted gene delivery to mammalian cells.
  • To enhance the efficiency of phage-mediated gene transfer through vector design and protocol modification.
  • To establish phage as a viable platform for gene therapy.

Main Methods:

  • Insertion of a mammalian reporter gene cassette (GFP) into phage vectors.
  • Fusion of phage pIII coat protein with cell-targeting ligands (FGF2, EGF).

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  • Modification of transfection protocols and vector design, including multivalent phagemid vectors.

    • Main Results:

    • Targeted phage gene delivery demonstrated concentration, time, and ligand dependency.
    • Phage particles showed specificity for target cell surface receptors.
    • Transduction efficiency was increased from 1-2% to up to 45% using optimized vectors and genotoxic treatment.

    Conclusions:

    • Genetic modification significantly enhances the efficiency of phage-mediated gene transfer.
    • Engineered phage vectors show potential for targeted gene delivery in mammalian cells.
    • Further evolution of phage vectors aims to improve targeting, stability, and reduce immunogenicity for gene therapy.