Improving AAV Vector Production for Gene Therapy
Author Information
Author(s): Shi Shuiliang, Mercer Scott A, Dilley Robert, Trippel Stephen B
Primary Institution: Indiana University School of Medicine
Hypothesis
Mutations in the start codon and upstream regulatory elements of Rep78/68 in AAV helper plasmids can regulate recombinant AAV vector production.
Conclusion
Selective mutations in Rep78/68 regulatory elements may enhance the therapeutic value of recombinant AAV vectors.
Supporting Evidence
- Mutations in the Rep78/68 start codon significantly affected rAAV production.
- The p5 promoter improved the function of the ACG start codon.
- The highest rAAV titer was achieved with the pAAV-RC/p5 construct.
Takeaway
Scientists are trying to make better tools for gene therapy by changing some parts of a virus to help it work better. This can help produce more of a helpful protein called IGF-I.
Methodology
AAV helper plasmids were constructed and co-transfected into 293 cells to produce rAAV-IGF-I, which was then used to transduce HT1080 cells.
Limitations
The study primarily focused on specific mutations and may not account for all factors affecting rAAV production.
Statistical Information
P-Value
p<0.001
Statistical Significance
p<0.001
Digital Object Identifier (DOI)
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