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Updated: Jun 12, 2026

Generation, Amplification, and Titration of Recombinant Respiratory Syncytial Viruses
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Published on: April 4, 2019

Titering replication-defective rSV40 vectors.

David S Strayer, Christine Mitchell, Dawn A Maier

    Cold Spring Harbor Protocols
    |June 3, 2010
    PubMed
    Summary
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    Recombinant simian virus 40 (rSV40) vectors offer efficient gene delivery to various cell types, including stem cells and neurons. Simple, reproducible methods ensure high-quality, replication-incompetent vectors for long-term gene expression.

    Area of Science:

    • Molecular Biology
    • Virology
    • Gene Therapy

    Background:

    • Recombinant simian virus 40 (rSV40)-derived vectors are effective for gene delivery.
    • These vectors target bone marrow progenitor cells, epithelial cells (e.g., hepatocytes), and central nervous system neurons and microglia.
    • rSV40 vectors integrate into cellular DNA for sustained gene expression in both resting and dividing cells, in vitro and in vivo.

    Purpose of the Study:

    • To describe simple, reproducible methods for producing, purifying, and quantifying rSV40 vectors.
    • To detail techniques for generating vectors with specific deletions (large T antigen or all coding sequences).
    • To present quantitative polymerase chain reaction (qPCR) as a method for titering and assessing purity of rSV40 vectors.

    Main Methods:

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    Last Updated: Jun 12, 2026

    Generation, Amplification, and Titration of Recombinant Respiratory Syncytial Viruses
    11:48

    Generation, Amplification, and Titration of Recombinant Respiratory Syncytial Viruses

    Published on: April 4, 2019

    Generation and Assembly of Virus-Specific Nucleocapsids of the Respiratory Syncytial Virus
    09:08

    Generation and Assembly of Virus-Specific Nucleocapsids of the Respiratory Syncytial Virus

    Published on: July 27, 2021

    Production of Replication-Defective Retrovirus by Transient Transfection of 293T cells
    09:40

    Production of Replication-Defective Retrovirus by Transient Transfection of 293T cells

    Published on: December 4, 2007

  • Vector production with deletions for large T antigen or all SV40 coding sequences, accommodating up to 5 kb of foreign DNA.
  • Purification via centrifugation using discontinuous sucrose or cesium chloride gradients.
  • Quantification and purity assessment using quantitative polymerase chain reaction (qPCR) with SYBR Green detection and melt curve analysis.

    • Main Results:

    • rSV40 vectors are replication-incompetent and free of wild-type SV40 revertants.
    • qPCR provides accurate titration of viral stocks and assessment of sample purity.
    • The described methods are simple, reproducible, and yield high-quality rSV40 vectors.

    Conclusions:

    • rSV40 vectors are versatile tools for gene delivery to diverse cell types.
    • Established purification and quantification methods ensure vector safety and efficacy.
    • Quantitative PCR is a reliable method for rSV40 vector characterization.