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SV40 as an effective gene transfer vector in vivo
1Department of Pathology, Anatomy and Cell Biology, Jefferson Medical College, Philadelphia, Pennsylvania 19107, USA.
The Journal of Biological Chemistry
|October 4, 1996
Summary
Replication-deficient simian virus 40 (SV40) carrying the luciferase gene successfully transferred genes in laboratory experiments and in mice. This gene transfer method showed no inflammatory response in vivo.
Area of Science:
- Molecular Biology
- Virology
- Gene Therapy
Background:
- Simian virus 40 (SV40) is a versatile tool for gene expression studies.
- Developing replication-deficient viral vectors is crucial for safe gene transfer applications.
Purpose of the Study:
- To engineer a replication-deficient recombinant SV40 vector for gene transfer.
- To evaluate the efficacy and safety of this vector for in vitro and in vivo gene delivery.
Main Methods:
- A recombinant SV40, named SVluc, was created by replacing the large T antigen gene with a polylinker and inserting the firefly luciferase gene.
- SVluc was transfected into Tag-expressing cells to produce a Tag-deficient, replication-deficient virus.
- In vitro transduction of luciferase expression was confirmed.
- BALB/c mice were inoculated with SVluc, and tissues were analyzed for luciferase expression and inflammatory responses from 3 to 21 days post-inoculation.
Main Results:
- SVluc successfully transduced functional luciferase expression in vitro.
- Luciferase protein was detected in mouse tissues from 3 to 21 days post-inoculation.
- No inflammatory reactions were observed in any tissues studied.
- Luciferase activity was measurable from 14 to 21 days post-inoculation.
Conclusions:
- Replication-deficient recombinant SV40 vectors can effectively mediate gene transfer both in vitro and in vivo.
- SVluc demonstrates a favorable safety profile with no observed inflammatory responses.
- This engineered SV40 vector holds potential for gene therapy applications.