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Dicistronic polioviruses as expression vectors for foreign genes

L Alexander1, H H Lu, M Gromeier

  • 1Department of Microbiology, School of Medicine, SUNY at Stony Brook 11794.

AIDS Research and Human Retroviruses
|January 1, 1994
PubMed
Summary

Researchers engineered novel polioviruses using internal ribosomal entry sites (IRES). These engineered viruses express foreign genes, including HIV-1 sequences, and elicit an immune response in transgenic mice.

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Area of Science:

  • Virology
  • Molecular Biology
  • Immunology

Background:

  • Picornaviruses utilize internal ribosomal entry sites (IRES) for cap-independent translation initiation.
  • Engineering viral genomes allows for the study of gene expression and the development of novel vectors.

Purpose of the Study:

  • To construct dicistronic polioviruses containing tandem IRES elements.
  • To insert foreign coding sequences into the poliovirus genome.
  • To evaluate the genetic stability and immunogenicity of engineered polioviruses expressing HIV-1 sequences.

Main Methods:

  • Construction of a novel viral RNA with tandem IRES elements (PV 5' NTR-EMCV IRES-PV ORF-3' NTR).
  • Transfection of RNA into HeLa cells to generate W1-PNENPO poliovirus.
  • Insertion of foreign open reading frames (ORFs) such as CAT, luciferase, or HIV-1 gp120 into the W1-PNENPO genome.

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  • Infection of transgenic mice expressing the poliovirus receptor (PVR+) with a dicistronic poliovirus (W1-PV/V3-3) containing HIV-1 sequences.

    • Main Results:

    • Viable dicistronic polioviruses were generated by inserting foreign coding sequences between tandem IRES elements.
    • Engineered poliovirus W1-PV/V3-3, containing HIV-1 V3 domain sequences, was successfully used to infect PVR+ transgenic mice.
    • The study discusses the genetic stability of these dicistronic viruses and the resulting HIV-1 specific immune response.

    Conclusions:

    • Novel dicistronic polioviruses can be engineered using tandem IRES elements to express foreign genes.
    • These engineered viruses demonstrate potential as vectors for studying gene expression and inducing immune responses.
    • Further investigation into the genetic stability and immune responses elicited by these novel viral constructs is warranted.