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Targeting vectors for intracellular immunisation

L Persic1, M Righi, A Roberts

  • 1Societa Italiana per la Ricerca Scientifica, Roma, Italy.

Gene
|March 10, 1997
PubMed
Summary
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Intracellular immunization uses engineered antibody fragments to inactivate specific molecules within cells. This novel approach allows researchers to study protein function in various cellular compartments using targeted recombinant antibodies.

Area of Science:

  • Molecular Biology
  • Immunology
  • Cell Biology

Background:

  • Intracellular immunization is a technique to inhibit molecular function within cells.
  • Recombinant antibodies can be targeted to specific cellular compartments.

Purpose of the Study:

  • To develop a versatile vector system for intracellular immunization.
  • To enable the study of molecules in diverse cellular locations.

Main Methods:

  • Modification of the VHExpress vector to express single-chain variable fragments (scFvs).
  • Inclusion of targeting signals for localization to endoplasmic reticulum, nucleus, mitochondria, cytoplasm, or secretion.
  • Assessment of vector function via immunofluorescence in transfected COS cells.

Main Results:

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  • Successfully created vectors enabling intracellular expression and targeting of scFvs.
  • Demonstrated localization of scFvs to various cellular compartments.
  • Validated the functionality of the developed intracellular immunization system.

Conclusions:

  • The developed vector system provides a powerful tool for intracellular immunization.
  • This technology facilitates the investigation of molecular functions across different cellular compartments.
  • Enables targeted inhibition and study of proteins within eukaryotic cells.