Academic Journal
A Robust System for Production of Superabundant VP1 Recombinant AAV Vectors
| Title: | A Robust System for Production of Superabundant VP1 Recombinant AAV Vectors |
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| Authors: | Qizhao Wang, Zhongren Wu, Junping Zhang, Jenni Firrman, Hongying Wei, Zhengjing Zhuang, LinShu Liu, Linqing Miao, Yang Hu, Dong Li, Yong Diao, Weidong Xiao |
| Source: | Molecular Therapy: Methods & Clinical Development, Vol 7, Iss C, Pp 146-156 (2017) |
| Publisher Information: | Elsevier, 2017. |
| Publication Year: | 2017 |
| Collection: | LCC:Genetics LCC:Cytology |
| Subject Terms: | AAV, vaccinia virus, VP1, gene therapy, vector production, Genetics, QH426-470, Cytology, QH573-671 |
| More Details: | 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 × 1015 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. |
| Document Type: | article |
| File Description: | electronic resource |
| Language: | English |
| ISSN: | 2329-0501 |
| Relation: | http://www.sciencedirect.com/science/article/pii/S2329050117301183; https://doaj.org/toc/2329-0501 |
| DOI: | 10.1016/j.omtm.2017.11.002 |
| Access URL: | https://doaj.org/article/af7b915df18d4bd5928b477858f894a5 |
| Accession Number: | edsdoj.f7b915df18d4bd5928b477858f894a5 |
| Database: | Directory of Open Access Journals |
| ISSN: | 23290501 |
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| DOI: | 10.1016/j.omtm.2017.11.002 |
| Published in: | Molecular Therapy: Methods & Clinical Development |
| Language: | English |