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A Robust System for Production of Superabundant VP1 Recombinant AAV Vectors
Recombinant adeno-associated viral (rAAV) vectors have been widely used in human gene therapy. One major impediment to its broad application is the inability to produce high-quality vectors in mass quantity. Here, an efficient and scalable suspension cell culture system for the production of rAAV ve...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society of Gene & Cell Therapy
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5721209/ https://www.ncbi.nlm.nih.gov/pubmed/29255740 http://dx.doi.org/10.1016/j.omtm.2017.11.002 |
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author | Wang, Qizhao Wu, Zhongren Zhang, Junping Firrman, Jenni Wei, Hongying Zhuang, Zhengjing Liu, LinShu Miao, Linqing Hu, Yang Li, Dong Diao, Yong Xiao, Weidong |
author_facet | Wang, Qizhao Wu, Zhongren Zhang, Junping Firrman, Jenni Wei, Hongying Zhuang, Zhengjing Liu, LinShu Miao, Linqing Hu, Yang Li, Dong Diao, Yong Xiao, Weidong |
author_sort | Wang, Qizhao |
collection | PubMed |
description | Recombinant adeno-associated viral (rAAV) vectors have been widely used in human gene therapy. One major impediment to its broad application is the inability to produce high-quality vectors in mass quantity. Here, an efficient and scalable suspension cell culture system for the production of rAAV vectors is described. In this system, the AAV trans factors, Rep78, Rep52, VP1, VP2, and VP3, were stably integrated into a single vaccinia virus carrier by maximizing the use of alternative codons between genes with identical amino acids, and the cis rAAV genome was carried by an E1/E3 gene-deleted adenovirus. Infection of improved, E1 integrated, suspension-cultured cells with these two viral vectors resulted in the robust production of rAAV vectors. The newly enhanced system can consistently produce ∼1 × 10(15) genome containing rAAV vectors per liter of suspension cells. Moreover, the capsid composition of rAAV vectors produced by this system is markedly different from those produced using the traditional system in that the VP1 protein is more abundant than the VP2 protein (19:1 versus 1:1). The unique VP1 superabundant rAAV vectors produced in this new system exhibited improved transduction in vivo after intravitreal injection. |
format | Online Article Text |
id | pubmed-5721209 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | American Society of Gene & Cell Therapy |
record_format | MEDLINE/PubMed |
spelling | pubmed-57212092017-12-18 A Robust System for Production of Superabundant VP1 Recombinant AAV Vectors Wang, Qizhao Wu, Zhongren Zhang, Junping Firrman, Jenni Wei, Hongying Zhuang, Zhengjing Liu, LinShu Miao, Linqing Hu, Yang Li, Dong Diao, Yong Xiao, Weidong Mol Ther Methods Clin Dev Article Recombinant adeno-associated viral (rAAV) vectors have been widely used in human gene therapy. One major impediment to its broad application is the inability to produce high-quality vectors in mass quantity. Here, an efficient and scalable suspension cell culture system for the production of rAAV vectors is described. In this system, the AAV trans factors, Rep78, Rep52, VP1, VP2, and VP3, were stably integrated into a single vaccinia virus carrier by maximizing the use of alternative codons between genes with identical amino acids, and the cis rAAV genome was carried by an E1/E3 gene-deleted adenovirus. Infection of improved, E1 integrated, suspension-cultured cells with these two viral vectors resulted in the robust production of rAAV vectors. The newly enhanced system can consistently produce ∼1 × 10(15) genome containing rAAV vectors per liter of suspension cells. Moreover, the capsid composition of rAAV vectors produced by this system is markedly different from those produced using the traditional system in that the VP1 protein is more abundant than the VP2 protein (19:1 versus 1:1). The unique VP1 superabundant rAAV vectors produced in this new system exhibited improved transduction in vivo after intravitreal injection. American Society of Gene & Cell Therapy 2017-11-07 /pmc/articles/PMC5721209/ /pubmed/29255740 http://dx.doi.org/10.1016/j.omtm.2017.11.002 Text en © 2017 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Article Wang, Qizhao Wu, Zhongren Zhang, Junping Firrman, Jenni Wei, Hongying Zhuang, Zhengjing Liu, LinShu Miao, Linqing Hu, Yang Li, Dong Diao, Yong Xiao, Weidong A Robust System for Production of Superabundant VP1 Recombinant AAV Vectors |
title | A Robust System for Production of Superabundant VP1 Recombinant AAV Vectors |
title_full | A Robust System for Production of Superabundant VP1 Recombinant AAV Vectors |
title_fullStr | A Robust System for Production of Superabundant VP1 Recombinant AAV Vectors |
title_full_unstemmed | A Robust System for Production of Superabundant VP1 Recombinant AAV Vectors |
title_short | A Robust System for Production of Superabundant VP1 Recombinant AAV Vectors |
title_sort | robust system for production of superabundant vp1 recombinant aav vectors |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5721209/ https://www.ncbi.nlm.nih.gov/pubmed/29255740 http://dx.doi.org/10.1016/j.omtm.2017.11.002 |
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