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Related Experiment Videos

Mammalian cell binding and transfection mediated by surface-modified bacteriophage lambda

I S Dunn1

  • 1Queensland Cancer Fund Research Unit, Department of Pathology, Medical School, University of Queensland Brisbane, Australia.

Biochimie
|January 1, 1996
PubMed
Summary
This summary is machine-generated.

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Bacteriophage lambda virions modified with Arg-Gly-Asp (RGD) peptides enhance mammalian cell adhesion and RGD-modified phage can efficiently transfect cells. This demonstrates that improving a single step in phage uptake significantly boosts transfection efficiency.

Area of Science:

  • * Molecular biology
  • * Virology
  • * Cell biology

Background:

  • * Bacteriophage lambda virions are viruses that infect bacteria.
  • * Cell surface receptors play a crucial role in cellular interactions and uptake processes.
  • * The Arg-Gly-Asp (RGD) peptide motif is known to bind to integrins on mammalian cells.

Purpose of the Study:

  • * To investigate the potential of modifying bacteriophage lambda virions with RGD peptides.
  • * To determine if RGD-modified phage can enhance mammalian cell adhesion to surfaces.
  • * To assess the efficacy of RGD-modified phage in mediating gene transfection into mammalian cells.

Main Methods:

  • * Genetic modification of bacteriophage lambda to incorporate cyclizable RGD peptides into major tail subunit (V) proteins.

Related Experiment Videos

  • * Cell adhesion assays using mammalian cells and modified/unmodified phage.
  • * Peptide competition experiments to confirm specificity of RGD-mediated binding.
  • * Transfection efficiency assays using COS cells and RGD-modified phage carrying a reporter gene.

    • Main Results:

    • * RGD-modified bacteriophage lambda virions significantly promoted the binding of mammalian cells to a solid surface.
    • * This cell-binding effect was specific and could be inhibited by competing peptides.
    • * Control phage lacking the RGD modification did not induce significant cellular interaction.
    • * RGD-modified phage, but not control phage, successfully transfected COS cells at a significant frequency.

    Conclusions:

    • * Modification of bacteriophage lambda with RGD peptides enhances mammalian cell adhesion.
    • * RGD-modified bacteriophage lambda can serve as an effective vehicle for gene delivery and transfection.
    • * Improving a single rate-limiting step, such as cell binding, can substantially increase the overall efficiency of phage-mediated transfection.