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Published on: April 8, 2012
Production of SV40-derived vectors
Cold Spring Harbor Protocols
|June 3, 2010
Summary
Recombinant simian virus 40 (rSV40)-derived vectors offer efficient, long-term gene delivery to various cell types. This protocol details simple, reproducible production and purification of these replication-incompetent vectors for diverse applications.
Area of Science:
- Molecular Biology
- Gene Therapy
- Virology
Background:
- Recombinant simian virus 40 (rSV40)-derived vectors are effective for gene delivery.
- These vectors target bone marrow progenitor cells, epithelial cells, and neurons.
- They enable rapid DNA integration for sustained gene expression in vitro and in vivo.
Purpose of the Study:
- To describe a protocol for producing and purifying rSV40-derived vectors.
- To enable gene delivery applications in various cell types.
- To facilitate the use of these vectors in research laboratories.
Main Methods:
- Utilizes packaging cells (e.g., COS-7) and circular vector genome DNA.
- Amplification via repeated infection of packaging cells with transfected vector.
- Purification by centrifugation using sucrose or cesium chloride (CsCl) gradients.
Main Results:
- Vectors are replication-incompetent with no detectable wild-type SV40 revertants.
- Protocols are simple, reproducible, and yield high-quality vectors.
- Vectors can carry up to 5 kb of foreign DNA, suitable for long-term expression.
Conclusions:
- The described methods simplify the production and purification of rSV40 vectors.
- These vectors are suitable for delivering mammalian or viral proteins and untranslated RNAs.
- The protocol allows broad laboratory access to rSV40 vector technology for gene delivery.
