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A packaging system for SV40 vectors without viral coding sequences

B Fang1, P Koch, M Bouvet

  • 1Department of Thoracic and Cardiovascular SurgeryJ, University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030, USA.

Analytical Biochemistry
|January 31, 1998
PubMed
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This study presents a novel system for packaging Simian virus 40 (SV40) vectors using recombinant adenoviruses. This method allows for efficient gene transfer vector production without viral coding sequences, yielding high-purity SV40 vectors.

Area of Science:

  • Molecular Biology
  • Virology
  • Gene Therapy

Background:

  • Simian virus 40 (SV40) vectors have been utilized for gene expression in mammalian cells since the 1980s.
  • Recent applications include gene transfer in mice and human peripheral blood cells.
  • Existing methods for SV40 vector production face limitations.

Purpose of the Study:

  • To develop an efficient system for packaging SV40 vectors.
  • To achieve high yields of infectious SV40 vectors free of viral coding sequences.
  • To optimize the SV40 vector packaging process using recombinant adenoviruses.

Main Methods:

  • A system was developed utilizing recombinant adenoviruses to express SV40 capsids.
  • Plasmids containing the SV40 replication origin were packaged.

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  • Helper adenoviruses were heat-inactivated post-packaging.

    • Main Results:

    • The system effectively packaged plasmids into SV40 capsids.
    • A high yield of infectious SV40 vector was achieved (approximately 3 x 10^5).
    • A high ratio of SV40 to adenoviral vector (approximately 1000:1) was obtained, with heat-inactivated helper adenoviruses showing no impact on SV40 vector infectivity.

    Conclusions:

    • A novel and efficient method for producing SV40 vectors has been established.
    • This system offers a high yield and purity of infectious SV40 vectors.
    • The developed packaging system is suitable for gene transfer applications.