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Viral Vector Production: Adenovirus

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Gene Therapy for Neurological Disorders

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1382))

Abstract

Adenoviral vectors have proven to be valuable resources in the development of novel therapies aimed at targeting pathological conditions of the central nervous system, including Alzheimer’s disease and neoplastic brain lesions. Not only can some genetically engineered adenoviral vectors achieve remarkably efficient and specific gene delivery to target cells, but they also may act as anticancer agents by selectively replicating within cancer cells.

Due to the great interest in using adenoviral vectors for various purposes, the need for a comprehensive protocol for viral vector production is especially apparent. Here, we describe the process of generating an adenoviral vector in its entirety, including the more complex process of adenoviral fiber modification to restrict viral tropism in order to achieve more efficient and specific gene delivery.

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References

  1. Majhen D, Ambriovic-Ristov A (2006) Adenoviral vectors—how to use them in cancer gene therapy? Virus Res 119:121–133

    Article  CAS  PubMed  Google Scholar 

  2. Green NK, Seymour LW (2002) Adenoviral vectors: systemic delivery and tumor targeting. Cancer Gene Ther 9:1036–1042

    Article  CAS  PubMed  Google Scholar 

  3. He TC et al (1998) A simplified system for generating recombinant adenoviruses. Proc Natl Acad Sci U S A 95:2509–2514

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  4. Graham FL (1987) Growth of 293 cells in suspension culture. J Gen Virol 68:937–940

    Article  PubMed  Google Scholar 

  5. Lavappa KS, Macy ML, Shannon JE (1976) Examination of ATCC stocks for HeLa marker chromosomes in human cell lines. Nature 259:211–213

    Article  CAS  PubMed  Google Scholar 

  6. Stulberg CS et al (1970) The animal cell culture collection. In Vitro 5:1–16

    Article  CAS  PubMed  Google Scholar 

  7. Graham FL et al (1977) Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol 36:59–74

    Article  CAS  PubMed  Google Scholar 

  8. Coecke S et al (2005) Guidance on good cell culture practice. A report of the second ECVAM task force on good cell culture practice. Altern Lab Anim 33:261–287

    CAS  PubMed  Google Scholar 

  9. Graham FL, Van der Eb AJ, Heijneker HL (1974) Size and location of the transforming region in human adenovirus type 5 DNA. Nature 251:687–691

    Article  CAS  PubMed  Google Scholar 

  10. Graham FL, Prevec L (1991) Manipulation of adenovirus vectors. Methods Mol Biol 7:109–128

    CAS  PubMed  Google Scholar 

  11. Sambrook J (2001) Molecular cloning: a laboratory manual. In: Russell DW (ed) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY

    Google Scholar 

  12. He TC (2004) Adenoviral vectors. Curr Protoc Hum Genet Chapter 12: Unit 12.4

    Google Scholar 

  13. Kim JW et al (2013) An adenovirus vector incorporating carbohydrate binding domains utilizes glycans for gene transfer. PLoS One 8:e55533

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  14. Hanahan D, Gluzman Y (1984) Rescue of functional replication origins from embedded configurations in a plasmid carrying the adenovirus genome. Mol Cell Biol 4:302–309

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  15. Imler JL et al (1995) An efficient procedure to select and recover recombinant adenovirus vectors. Gene Ther 2:263–268

    CAS  PubMed  Google Scholar 

  16. Chartier C et al (1996) Efficient generation of recombinant adenovirus vectors by homologous recombination in Escherichia coli. J Virol 70:4805–4810

    PubMed Central  CAS  PubMed  Google Scholar 

  17. Krougliak V, Graham FL (1995) Development of cell lines capable of complementing E1, E4, and protein IX defective adenovirus type 5 mutants. Hum Gene Ther 6:1575–1586

    Article  CAS  PubMed  Google Scholar 

  18. Ng P, Graham FL (2002) Construction of first-generation adenoviral vectors. Methods Mol Med 69:389–414

    CAS  PubMed  Google Scholar 

  19. Kanegae Y, Makimura M, Saito I (1994) A simple and efficient method for purification of infectious recombinant adenovirus. Jpn J Med Sci Biol 47:157–166

    Article  CAS  PubMed  Google Scholar 

  20. Maizel JV, White DO Jr, Scharff MD (1968) The polypeptides of adenovirus. I. Evidence for multiple protein components in the virion and a comparison of types 2, 7A, and 12. Virology 36:115–125

    Article  CAS  PubMed  Google Scholar 

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Correspondence to Maciej S. Lesniak .

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Kim, J.W., Morshed, R.A., Kane, J.R., Auffinger, B., Qiao, J., Lesniak, M.S. (2016). Viral Vector Production: Adenovirus. In: Manfredsson, F. (eds) Gene Therapy for Neurological Disorders. Methods in Molecular Biology, vol 1382. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3271-9_9

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  • DOI: https://doi.org/10.1007/978-1-4939-3271-9_9

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3270-2

  • Online ISBN: 978-1-4939-3271-9

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