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Adenovirus-Mediated Gene Transfer

  • Ko Willems van Dijk
  • Kyriakos E. Kypreos
  • Frits J. Fallaux
  • Jurre HagemanEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 693)

Abstract

Recombinant adenoviruses are attractive vectors for short-term expression in mouse liver and primary cell lines. Various versatile vector systems have been developed which can be used for the reliable production of recombinant adenoviruses. This protocol describes the entire process for the production of recombinant adenoviruses using the AdEasy system. This protocol will give a practical step-by-step description from the cloning of the gene of interest until the in vivo administration in mice. The entire process will take about 8 weeks to complete.

Key words

Recombinant adenoviruses Adenovirus AdEasy pAdEasy Viral transduction Transduction 

References

  1. 1.
    Shen, C. and Reske, S. N. (2004) Adenovirus-delivered siRNA, Methods Mol. Biol. 252, 523–532.PubMedGoogle Scholar
  2. 2.
    Hosono, T., Mizuguchi, H., Katayama, K., Xu, Z. L., Sakurai, F., Ishii-Watabe, A., Kawabata, K., Yamaguchi, T., Nakagawa, S., Mayumi, T., and Hayakawa, T. (2004) Adenovirus vector-mediated doxycycline-inducible RNA interference, Hum. Gene Ther. 15, 813–819.PubMedCrossRefGoogle Scholar
  3. 3.
    Gomez-Valades, A. G., Mendez-Lucas, A., Vidal-Alabro, A., Blasco, F. X., Chillon, M., Bartrons, R., Bermudez, J., and Perales, J. C. (2008) Pck1 gene silencing in the liver improves glycemia control, insulin sensitivity, and dyslipidemia in db/db mice, Diabetes 57, 2199–2210.PubMedCrossRefGoogle Scholar
  4. 4.
    Russell, W. C. (2000) Update on adenovirus and its vectors, J. Gen. Virol. 81, 2573–2604.PubMedGoogle Scholar
  5. 5.
    McGrory, W. J., Bautista, D. S., and Graham, F. L. (1988) A simple technique for the rescue of early region I mutations into infectious human adenovirus type 5, Virology 163, 614–617.PubMedCrossRefGoogle Scholar
  6. 6.
    He, T. C., Zhou, S., da Costa, L. T., Yu, J., Kinzler, K. W., and Vogelstein, B. (1998) A simplified system for generating recombinant adenoviruses, Proc. Natl. Acad. Sci. U.S.A. 95, 2509–2514.PubMedCrossRefGoogle Scholar
  7. 7.
    Fu, S. and Deisseroth, A. B. (1997) Use of the cosmid adenoviral vector cloning system for the in vitro construction of recombinant adenoviral vectors, Hum. Gene Ther. 8, 1321–1330.PubMedCrossRefGoogle Scholar
  8. 8.
    Ng, P., Parks, R. J., Cummings, D. T., Evelegh, C. M., Sankar, U., and Graham, F. L. (1999) A high-efficiency Cre/loxP-based system for construction of adenoviral vectors, Hum. Gene Ther. 10, 2667–2672.PubMedCrossRefGoogle Scholar
  9. 9.
    Ng, P., Cummings, D. T., Evelegh, C. M., and Graham, F. L. (2000) Yeast recombinase FLP functions effectively in human cells for construction of adenovirus vectors, BioTechniques 29, 524–526, 528.Google Scholar
  10. 10.
    Tashiro, F., Niwa, H., and Miyazaki, J. (1999) Constructing adenoviral vectors by using the circular form of the adenoviral genome cloned in a cosmid and the Cre-loxP recombination system, Hum. Gene Ther. 10, 1845–1852.PubMedCrossRefGoogle Scholar
  11. 11.
    Ng, P., Parks, R. J., Cummings, D. T., Evelegh, C. M., and Graham, F. L. (2000) An enhanced system for construction of adenoviral vectors by the two-plasmid rescue method, Hum. Gene Ther. 11, 693–699.PubMedCrossRefGoogle Scholar
  12. 12.
    Aoki, K., Barker, C., Danthinne, X., Imperiale, M. J., and Nabel, G. J. (1999) Efficient generation of recombinant adenoviral vectors by Cre-lox recombination in vitro, Mol. Med. 5, 224–231.PubMedGoogle Scholar
  13. 13.
    Luo, J., Deng, Z. L., Luo, X., Tang, N., Song, W. X., Chen, J., Sharff, K. A., Luu, H. H., Haydon, R. C., Kinzler, K. W., Vogelstein, B., and He, T. C. (2007) A protocol for rapid generation of recombinant adenoviruses using the AdEasy system, Nat. Protoc. 2, 1236–1247.PubMedCrossRefGoogle Scholar
  14. 14.
    Fallaux, F. J., Kranenburg, O., Cramer, S. J., Houweling, A., Van, O. H., Hoeben, R. C., and Van Der Eb, A. J. (1996) Characterization of 911: a new helper cell line for the titration and propagation of early region 1-deleted adenoviral vectors, Hum. Gene Ther. 7, 215–222.PubMedCrossRefGoogle Scholar
  15. 15.
    Fallaux, F. J., Bout, A., van der Velde, I., van den Wollenberg, D. J., Hehir, K. M., Keegan, J., Auger, C., Cramer, S. J., Van, O. H., Van Der Eb, A. J., Valerio, D., and Hoeben, R. C. (1998) New helper cells and matched early region 1-deleted adenovirus vectors prevent generation of replication-competent adenoviruses, Hum. Gene Ther. 9, 1909–1917.PubMedCrossRefGoogle Scholar
  16. 16.
    Berdichevsky, M., Gentile, M. P., Hughes, B., Meis, P., Peltier, J., Blumentals, I., Aunins, J., and Altaras, N. E. (2008) Establishment of higher passage PER.C6 cells for adenovirus manufacture, Biotechnol. Prog. 24, 158–165.PubMedCrossRefGoogle Scholar
  17. 17.
    Schagen, F. H., Rademaker, H. J., Rabelink, M. J., Van, O. H., Fallaux, F. J., Van Der Eb, A. J., and Hoeben, R. C. (2000) Ammonium sulphate precipitation of recombinant adenovirus from culture medium: an easy method to increase the total virus yield, Gene Ther. 7, 1570–1574.PubMedCrossRefGoogle Scholar
  18. 18.
    Fallaux, F. J., Van Der Eb, A. J., and Hoeben, R. C. (1999) Who’s afraid of replication-competent adenoviruses? Gene Ther. 6, 709–712.PubMedCrossRefGoogle Scholar
  19. 19.
    Pietersen, A. M., van der Eb, M. M., Rademaker, H. J., van den Wollenberg, D. J., Rabelink, M. J., Kuppen, P. J., van Dierendonck, J. H., Van, O. H., Masman, D., van de Velde, C. J., van der Eb, A. J., Hoeben, R. C., and Noteborn, M. H. (1999) Specific tumor-cell killing with adenovirus vectors containing the apoptin gene, Gene Ther. 6, 882–892.PubMedCrossRefGoogle Scholar
  20. 20.
    Yang, Y., Xiang, Z., Ertl, H. C., and Wilson, J. M. (1995) Upregulation of class I major histocompatibility complex antigens by interferon gamma is necessary for T-cell-mediated elimination of recombinant adenovirus-infected hepatocytes in vivo, Proc. Natl. Acad. Sci. U.S.A. 92, 7257–7261.PubMedCrossRefGoogle Scholar
  21. 21.
    Steel, C. D., Stephens, A. L., Hahto, S. M., Singletary, S. J., and Ciavarra, R. P. (2008) Comparison of the lateral tail vein and the retro-orbital venous sinus as routes of intravenous drug delivery in a transgenic mouse model, Lab Anim (NY) 37, 26–32.CrossRefGoogle Scholar
  22. 22.
    Kay, M. A., Holterman, A. X., Meuse, L., Gown, A., Ochs, H. D., Linsley, P. S., and Wilson, C. B. (1995) Long-term hepatic adenovirus-mediated gene expression in mice following CTLA4Ig administration, Nat. Genet. 11, 191–197.PubMedCrossRefGoogle Scholar
  23. 23.
    Jooss, K., Yang, Y., and Wilson, J. M. (1996) Cyclophosphamide diminishes inflammation and prolongs transgene expression following delivery of adenoviral vectors to mouse liver and lung, Hum. Gene Ther. 7, 1555–1566.PubMedCrossRefGoogle Scholar
  24. 24.
    Engelhardt, J. F., Litzky, L., and Wilson, J. M. (1994) Prolonged transgene expression in cotton rat lung with recombinant adenoviruses defective in E2a, Hum. Gene Ther. 5, 1217–1229.PubMedCrossRefGoogle Scholar
  25. 25.
    Yang, Y., Nunes, F. A., Berencsi, K., Gonczol, E., Engelhardt, J. F., and Wilson, J. M. (1994) Inactivation of E2a in recombinant adenoviruses improves the prospect for gene therapy in cystic fibrosis, Nat. Genet. 7, 362–369.PubMedCrossRefGoogle Scholar
  26. 26.
    Yeh, P., Dedieu, J. F., Orsini, C., Vigne, E., Denefle, P., and Perricaudet, M. (1996) Efficient dual transcomplementation of adenovirus E1 and E4 regions from a 293-derived cell line expressing a minimal E4 functional unit, J. Virol. 70, 559–565.PubMedGoogle Scholar
  27. 27.
    Wang, Q., Greenburg, G., Bunch, D., Farson, D., and Finer, M. H. (1997) Persistent transgene expression in mouse liver following in vivo gene transfer with a delta E1/delta E4 adenovirus vector, Gene Ther. 4, 393–400.PubMedCrossRefGoogle Scholar
  28. 28.
    Andrews, J. L., Kadan, M. J., Gorziglia, M. I., Kaleko, M., and Connelly, S. (2001) Generation and characterization of E1/E2a/E3/E4-deficient adenoviral vectors encoding human factor VIII, Mol. Ther. 3, 329–336.PubMedCrossRefGoogle Scholar
  29. 29.
    Parks, R. J., Chen, L., Anton, M., Sankar, U., Rudnicki, M. A., and Graham, F. L. (1996) A helper-dependent adenovirus vector system: removal of helper virus by Cre-mediated excision of the viral packaging signal, Proc. Natl. Acad. Sci. U.S.A. 93, 13565–13570.PubMedCrossRefGoogle Scholar
  30. 30.
    Ng, P., Beauchamp, C., Evelegh, C., Parks, R., and Graham, F. L. (2001) Development of a FLP/frt system for generating helper-dependent adenoviral vectors, Mol. Ther. 3, 809–815.PubMedCrossRefGoogle Scholar
  31. 31.
    Alba, R., Hearing, P., Bosch, A., and Chillon, M. (2007) Differential amplification of adenovirus vectors by flanking the packaging signal with attB/attP-PhiC31 sequences: implications for helper-dependent adenovirus production, Virology 367, 51–58.PubMedCrossRefGoogle Scholar
  32. 32.
    Morsy, M. A., Gu, M., Motzel, S., Zhao, J., Lin, J., Su, Q., Allen, H., Franlin, L., Parks, R. J., Graham, F. L., Kochanek, S., Bett, A. J., and Caskey, C. T. (1998) An adenoviral vector deleted for all viral coding sequences results in enhanced safety and extended expression of a leptin transgene, Proc. Natl. Acad. Sci. U.S.A. 95, 7866–7871.PubMedCrossRefGoogle Scholar
  33. 33.
    McCarty, D. M., Fu, H., Monahan, P. E., Toulson, C. E., Naik, P., and Samulski, R. J. (2003) Adeno-associated virus terminal repeat (TR) mutant generates self-complementary vectors to overcome the rate-limiting step to transduction in vivo, Gene Ther. 10, 2112–2118.PubMedCrossRefGoogle Scholar
  34. 34.
    Gao, G. P., Lu, Y., Sun, X., Johnston, J., Calcedo, R., Grant, R., and Wilson, J. M. (2006) High-level transgene expression in nonhuman primate liver with novel adeno-associated virus serotypes containing self-complementary genomes, J. Virol. 80, 6192–6194.PubMedCrossRefGoogle Scholar
  35. 35.
    Greber, U. F., Willetts, M., Webster, P., and Helenius, A. (1993) Stepwise dismantling of adenovirus 2 during entry into cells, Cell 75, 477–486.PubMedCrossRefGoogle Scholar
  36. 36.
    Nakano, M. Y., Boucke, K., Suomalainen, M., Stidwill, R. P., and Greber, U. F. (2000) The first step of adenovirus type 2 disassembly occurs at the cell surface, independently of endocytosis and escape to the cytosol, J. Virol. 74, 7085–7095.PubMedCrossRefGoogle Scholar
  37. 37.
    Bergelson, J. M., Cunningham, J. A., Droguett, G., Kurt-Jones, E. A., Krithivas, A., Hong, J. S., Horwitz, M. S., Crowell, R. L., and Finberg, R. W. (1997) Isolation of a common receptor for Coxsackie B viruses and adenoviruses 2 and 5, Science 275, 1320–1323.PubMedCrossRefGoogle Scholar
  38. 38.
    Krasnykh, V., Dmitriev, I., Mikheeva, G., Miller, C. R., Belousova, N., and Curiel, D. T. (1998) Characterization of an adenovirus vector containing a heterologous peptide epitope in the HI loop of the fiber knob, J. Virol. 72, 1844–1852.PubMedGoogle Scholar
  39. 39.
    Michael, S. I., Hong, J. S., Curiel, D. T., and Engler, J. A. (1995) Addition of a short peptide ligand to the adenovirus fiber protein, Gene Ther. 2, 660–668.PubMedGoogle Scholar
  40. 40.
    Wickham, T. J., Roelvink, P. W., Brough, D. E., and Kovesdi, I. (1996) Adenovirus targeted to heparan-containing receptors increases its gene delivery efficiency to multiple cell types, Nat. Biotechnol. 14, 1570–1573.PubMedCrossRefGoogle Scholar
  41. 41.
    Vigne, E., Mahfouz, I., Dedieu, J. F., Brie, A., Perricaudet, M., and Yeh, P. (1999) RGD inclusion in the hexon monomer provides adenovirus type 5-based vectors with a fiber knob-independent pathway for infection, J. Virol. 73, 5156–5161.PubMedGoogle Scholar
  42. 42.
    Miller, C. R., Buchsbaum, D. J., Reynolds, P. N., Douglas, J. T., Gillespie, G. Y., Mayo, M. S., Raben, D., and Curiel, D. T. (1998) Differential susceptibility of primary and established human glioma cells to adenovirus infection: targeting via the epidermal growth factor receptor achieves fiber receptor-independent gene transfer, Cancer Res. 58, 5738–5748.PubMedGoogle Scholar
  43. 43.
    Douglas, J. T., Rogers, B. E., Rosenfeld, M. E., Michael, S. I., Feng, M., and Curiel, D. T. (1996) Targeted gene delivery by tropism-modified adenoviral vectors, Nat. Biotechnol. 14, 1574–1578.PubMedCrossRefGoogle Scholar
  44. 44.
    Wickham, T. J., Segal, D. M., Roelvink, P. W., Carrion, M. E., Lizonova, A., Lee, G. M., and Kovesdi, I. (1996) Targeted adenovirus gene transfer to endothelial and smooth muscle cells by using bispecific antibodies, J. Virol. 70, 6831–6838.PubMedGoogle Scholar
  45. 45.
    Stevenson, S. C., Rollence, M., White, B., Weaver, L., and McClelland, A. (1995) Human adenovirus serotypes 3 and 5 bind to two different cellular receptors via the fiber head domain, J. Virol. 69, 2850–2857.PubMedGoogle Scholar
  46. 46.
    Krasnykh, V. N., Mikheeva, G. V., Douglas, J. T., and Curiel, D. T. (1996) Generation of recombinant adenovirus vectors with modified fibers for altering viral tropism, J. Virol. 70, 6839–6846.PubMedGoogle Scholar
  47. 47.
    Gall, J., Kass-Eisler, A., Leinwand, L., and Falck-Pedersen, E. (1996) Adenovirus type 5 and 7 capsid chimera: fiber replacement alters receptor tropism without affecting primary immune neutralization epitopes, J. Virol. 70, 2116–2123.PubMedGoogle Scholar
  48. 48.
    Wickham, T. J., Tzeng, E., Shears, L. L., Roelvink, P. W., Li, Y., Lee, G. M., Brough, D. E., Lizonova, A., and Kovesdi, I. (1997) Increased in vitro and in vivo gene transfer by adenovirus vectors containing chimeric fiber proteins, J. Virol. 71, 8221–8229.PubMedGoogle Scholar
  49. 49.
    Arnberg, N. (2009) Adenovirus receptors: implications for tropism, treatment and targeting, Rev. Med. Virol. 19, 165–178.PubMedCrossRefGoogle Scholar
  50. 50.
    Schiedner, G., Hertel, S., and Kochanek, S. (2000) Efficient transformation of primary human amniocytes by E1 functions of Ad5: generation of new cell lines for adenoviral vector production, Hum. Gene Ther. 11, 2105–2116.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Ko Willems van Dijk
    • 1
  • Kyriakos E. Kypreos
    • 2
  • Frits J. Fallaux
    • 3
  • Jurre Hageman
    • 4
    Email author
  1. 1.Departments of Human and Clinical GeneticsLeiden University Medical CenterLeidenThe Netherlands
  2. 2.Pharmacology Unit, Department of MedicineUniversity of Patras Medical SchoolRioGreece
  3. 3.Utrecht HoldingsUniversity of UtrechtUtrechtThe Netherlands
  4. 4.Laboratory of Pediatrics, Digestive and Metabolic Diseases Center for LiverUniversity Medical Center Groningen, University of GroningenGroningenThe Netherlands

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