Adeno-associated virus vectors can be efficiently produced without helper virus - PubMed (original) (raw)
Adeno-associated virus vectors can be efficiently produced without helper virus
T Matsushita et al. Gene Ther. 1998 Jul.
Abstract
The purpose of this work was to develop an efficient method for the production of adeno-associated virus (AAV) vectors in the absence of helper virus. The adenovirus regions that mediate AAV vector replication were identified and assembled into a helper plasmid. These included the VA, E2A and E4 regions. When this helper plasmid was cotransfected into 293 cells, along with plasmids encoding the AAV vector, and rep and cap genes, AAV vector was produced as efficiently as when using adenovirus infection as a source of help. CMV-driven constructs expressing the E4orf6 and the 72-M(r), E2A proteins were able to functionally replace the E4 and E2A regions, respectively. Therefore the minimum set of genes required to produce AAV helper activity equivalent to that provided by adenovirus infection consists of, or is a subset of, the following genes: the E4orf6 gene, the 72-M(r), E2A protein gene, the VA RNA genes and the E1 region. AAV vector preparations made with adenovirus and by the helper virus-free method were essentially indistinguishable with respect to particle density, particle to infectivity ratio, capsimer ratio and efficiency of muscle transduction in vivo. Only AAV vector preparations made by the helper virus-free method were not reactive with anti-adenovirus sera.
Similar articles
- An Improved Helper Plasmid Containing Deletions Within the E4 and E2a Genes Results in Increased Adeno-Associated Virus Productivity.
van Lieshout L, Ota S, Adusei A, Wiberg E, Costa-Grant K, Lata D, Dollive S, Stanvick M, Iwuchukwu I, Golebiowski D, Yin J. van Lieshout L, et al. Hum Gene Ther. 2024 Sep;35(17-18):767-776. doi: 10.1089/hum.2024.059. Epub 2024 Jul 31. Hum Gene Ther. 2024. PMID: 39001832 - Improved adeno-associated virus vector production with transfection of a single helper adenovirus gene, E4orf6.
Allen JM, Halbert CL, Miller AD. Allen JM, et al. Mol Ther. 2000 Jan;1(1):88-95. doi: 10.1006/mthe.1999.0010. Mol Ther. 2000. PMID: 10933916 - A helper virus-free packaging system for recombinant adeno-associated virus vectors.
Collaco RF, Cao X, Trempe JP. Collaco RF, et al. Gene. 1999 Oct 1;238(2):397-405. doi: 10.1016/s0378-1119(99)00347-9. Gene. 1999. PMID: 10570967 - The Interplay between Adeno-Associated Virus and its Helper Viruses.
Meier AF, Fraefel C, Seyffert M. Meier AF, et al. Viruses. 2020 Jun 19;12(6):662. doi: 10.3390/v12060662. Viruses. 2020. PMID: 32575422 Free PMC article. Review. - Helper virus-free gutless adenovirus (HF-GLAd): a new platform for gene therapy.
Liu J, Seol DW. Liu J, et al. BMB Rep. 2020 Nov;53(11):565-575. doi: 10.5483/BMBRep.2020.53.11.185. BMB Rep. 2020. PMID: 32958121 Free PMC article. Review.
Cited by
- Synergism of dual AAV gene therapy and rapamycin rescues GSDIII phenotype in muscle and liver.
Jauze L, Vie M, Miagoux Q, Rossiaud L, Vidal P, Montalvo-Romeral V, Saliba H, Jarrige M, Polveche H, Nozi J, Le Brun PR, Bocchialini L, Francois A, Cosette J, Rouillon J, Collaud F, Bordier F, Bertil-Froidevaux E, Georger C, van Wittenberghe L, Miranda A, Daniele NF, Gross DA, Hoch L, Nissan X, Ronzitti G. Jauze L, et al. JCI Insight. 2024 May 16;9(11):e172614. doi: 10.1172/jci.insight.172614. JCI Insight. 2024. PMID: 38753465 Free PMC article. - Engineering a synthetic gene circuit for high-performance inducible expression in mammalian systems.
De Carluccio G, Fusco V, di Bernardo D. De Carluccio G, et al. Nat Commun. 2024 Apr 17;15(1):3311. doi: 10.1038/s41467-024-47592-y. Nat Commun. 2024. PMID: 38632224 Free PMC article. - Adeno-associated virus as a delivery vector for gene therapy of human diseases.
Wang JH, Gessler DJ, Zhan W, Gallagher TL, Gao G. Wang JH, et al. Signal Transduct Target Ther. 2024 Apr 3;9(1):78. doi: 10.1038/s41392-024-01780-w. Signal Transduct Target Ther. 2024. PMID: 38565561 Free PMC article. Review. - Development of Stable Packaging and Producer Cell Lines for the Production of AAV Vectors.
Merten OW. Merten OW. Microorganisms. 2024 Feb 13;12(2):384. doi: 10.3390/microorganisms12020384. Microorganisms. 2024. PMID: 38399788 Free PMC article. Review. - Recent advances in various adeno-associated viruses (AAVs) as gene therapy agents in hepatocellular carcinoma.
Hadi M, Qutaiba B Allela O, Jabari M, Jasoor AM, Naderloo O, Yasamineh S, Gholizadeh O, Kalantari L. Hadi M, et al. Virol J. 2024 Jan 12;21(1):17. doi: 10.1186/s12985-024-02286-1. Virol J. 2024. PMID: 38216938 Free PMC article. Review.
MeSH terms
LinkOut - more resources
Full Text Sources
Other Literature Sources
Miscellaneous