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Cloning and Large-Scale Production of High-Capacity Adenoviral Vectors Based on the Human Adenovirus Type 5
Published on: January 28, 2016
The herpes simplex virus amplicon: a new eucaryotic defective-virus cloning-amplifying vector
Cell
|August 1, 1982
Summary
Researchers developed a herpes simplex virus (HSV) amplicon system for cloning and amplifying DNA in eukaryotic cells. This novel vector system efficiently replicates and can be propagated in virus stocks, enabling gene transfer applications.
Area of Science:
- Molecular Biology
- Virology
- Gene Therapy
Background:
- Herpes simplex virus (HSV) defective genomes can be regenerated into concatemeric forms.
- Previous studies showed cotransfection of cells with helper virus DNA and HSV defective genomes leads to regeneration.
Purpose of the Study:
- To derive a cloning-amplifying vector (amplicon) based on HSV defective genomes.
- To enable replication in eukaryotic cells with the assistance of a standard HSV helper virus.
Main Methods:
- Employing repeat units of HSV defective genomes to create an amplicon vector.
- Cotransfecting cells with helper virus DNA and chimeric repeat units (HSV-pKC7 DNA).
- Packaging chimeric defective genomes into virus particles for propagation.
Main Results:
- Generation of defective genomes composed of HSV-pKC7 repeat reiterations.
- Successful packaging and propagation of chimeric defective genomes in virus stocks, reaching over 90% enrichment.
- Demonstrated bidirectional transfer of monomeric chimeric repeat units between bacterial and eukaryotic cells.
Conclusions:
- The HSV amplicon system is a viable tool for cloning and amplification in eukaryotic cells.
- The system allows for efficient propagation and potential for gene delivery applications.
- The vector's design facilitates the introduction of additional DNA sequences into eukaryotic cells.
