DOI: 10.1073/pnas.97.3.1002. Optimization of the helper-dependent adenovirus system for production and potency <i>in</i> <i>vivo</i>

Optimization of the helper-dependent adenovirus system for production and potency in vivo

10.1073/pnas.97.3.1002

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Helper-dependent (HD) adenoviral vectors devoid of all viral coding sequences provide for safe and highly efficient gene transfer with long-lasting transgene expression. High titer stocks of HD vectors can be generated by using the cre-recombinase system. However, we have encountered difficulties with this system, including rearranged HD vectors and variable efficiency of HD vector rescue. These problems represent a major hindrance, particularly with regard to large-scale production. To overcome these limitations, we have modified the system in two ways: We constructed a new helper virus with a modified packaging signal and enhanced growth characteristics. We also redesigned the vector backbones by including noncoding adenovirus sequences adjacent to the right inverted terminal repeat and by incorporated a number of different segments of noncoding DNA of human origin as “stuffer.” Comparison of these vectors showed that the nature of the stuffer sequence affects replication of the HD vector. Optimization of the system resulted in a more robust and consistent production of HD vectors with low helper contamination and high in vivo potency.

Bibliography

Sandig, V., Youil, R., Bett, A. J., Franlin, L. L., Oshima, M., Maione, D., Wang, F., Metzker, M. L., Savino, R., & Caskey, C. T. (2000). Optimization of the helper-dependent adenovirus system for production and potency in vivo. Proceedings of the National Academy of Sciences, 97(3), 1002â1007.

Authors 10

Volker Sandig (first)
Rima Youil (additional)
Andrew J. Bett (additional)
Laura L. Franlin (additional)
Masanobu Oshima (additional)
Domenico Maione (additional)
Fubao Wang (additional)
Michael L. Metzker (additional)
Rocco Savino (additional)
C. Thomas Caskey (additional)

References 46 Referenced 171

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Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:32 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 5:49 p.m.)
Indexed	1 month, 2 weeks ago (July 14, 2025, 11:20 p.m.)
Issued	25 years, 7 months ago (Feb. 1, 2000)
Published	25 years, 7 months ago (Feb. 1, 2000)
Published Online	25 years, 7 months ago (Feb. 1, 2000)
Published Print	25 years, 7 months ago (Feb. 1, 2000)

Funders 0

None

BibTeX

@article{Sandig_2000, title={Optimization of the helper-dependent adenovirus system for production and potency in vivo}, volume={97}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.97.3.1002}, DOI={10.1073/pnas.97.3.1002}, number={3}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Sandig, Volker and Youil, Rima and Bett, Andrew J. and Franlin, Laura L. and Oshima, Masanobu and Maione, Domenico and Wang, Fubao and Metzker, Michael L. and Savino, Rocco and Caskey, C. Thomas}, year={2000}, month=feb, pages={1002–1007} }

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