Impact of Vector Design and Host Cell on Retroviral Vector Mutations
Author Information
Author(s): Paar Matthias, Klein Dieter, Salmons Brian, Günzburg Walter H, Renner Matthias, Portsmouth Daniel
Primary Institution: University of Veterinary Medicine, Vienna, Austria
Hypothesis
How do vector design and host cell type affect the emergence of mutant subpopulations in replicating retroviral vectors?
Conclusion
MLV-based RCR vectors with the transgene cassette in the 3' UTR are more stable and effective than those with the cassette in the U3 region.
Supporting Evidence
- MLV-based RCR vectors showed different rates of mutant emergence depending on the host cell type.
- Vectors with the transgene in the 3' UTR were more stable than those with the transgene in the U3 region.
- Real-time PCR was used to quantify the emergence of vector recombinants in both in vitro and in vivo settings.
Takeaway
This study shows that the design of a virus and the type of cell it infects can change how many mutant viruses are created, which is important for gene therapy.
Methodology
The study involved real-time PCR and RT-PCR to analyze vector genomes and proviral DNA from infected cells and tumors.
Potential Biases
Potential bias due to the use of specific cell lines and vector strains.
Limitations
The study primarily focused on specific vector designs and may not generalize to all retroviral vectors.
Participant Demographics
In vivo studies were conducted using nude mice with subcutaneous tumors.
Digital Object Identifier (DOI)
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