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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1994 Sep 13;91(19):8915–8919. doi: 10.1073/pnas.91.19.8915

Adeno-associated virus vectors preferentially transduce cells in S phase.

D W Russell 1, A D Miller 1, I E Alexander 1
PMCID: PMC44717  PMID: 8090744

Abstract

Vectors based on adeno-associated virus can stably transfer genes by chromosomal integration in recipient cells. In this study we have infected stationary and dividing primary human fibroblast cultures with adeno-associated virus vectors encoding alkaline phosphatase and neomycin phosphotransferase. We find that the transduction frequency of S phase cells is about 200 times that of non-S phase cells. However, neither S phase nor mitosis is essential for transduction. Single-stranded vector genomes survive in stationary cultures and can be recruited for transduction by stimulating these cultures to divide. Stable transductants contain randomly integrated vector sequences. These findings have important implications for the use of adeno-associated virus vectors in gene therapy.

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Selected References

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  1. Berns K. I., Pinkerton T. C., Thomas G. F., Hoggan M. D. Detection of adeno-associated virus (AAV)-specific nucleotide sequences in DNA isolated from latently infected Detroit 6 cells. Virology. 1975 Dec;68(2):556–560. doi: 10.1016/0042-6822(75)90298-6. [DOI] [PubMed] [Google Scholar]
  2. Fields-Berry S. C., Halliday A. L., Cepko C. L. A recombinant retrovirus encoding alkaline phosphatase confirms clonal boundary assignment in lineage analysis of murine retina. Proc Natl Acad Sci U S A. 1992 Jan 15;89(2):693–697. doi: 10.1073/pnas.89.2.693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Flotte T. R., Solow R., Owens R. A., Afione S., Zeitlin P. L., Carter B. J. Gene expression from adeno-associated virus vectors in airway epithelial cells. Am J Respir Cell Mol Biol. 1992 Sep;7(3):349–356. doi: 10.1165/ajrcmb/7.3.349. [DOI] [PubMed] [Google Scholar]
  4. Graham F. L., Smiley J., Russell W. C., Nairn R. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol. 1977 Jul;36(1):59–74. doi: 10.1099/0022-1317-36-1-59. [DOI] [PubMed] [Google Scholar]
  5. Handa H., Shiroki K., Shimojo H. Establishment and characterization of KB cell lines latently infected with adeno-associated virus type 1. Virology. 1977 Oct 1;82(1):84–92. doi: 10.1016/0042-6822(77)90034-4. [DOI] [PubMed] [Google Scholar]
  6. Hermonat P. L., Muzyczka N. Use of adeno-associated virus as a mammalian DNA cloning vector: transduction of neomycin resistance into mammalian tissue culture cells. Proc Natl Acad Sci U S A. 1984 Oct;81(20):6466–6470. doi: 10.1073/pnas.81.20.6466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  8. Kollek R., Goulian M. Synthesis of parvovirus H-1 replicative form from viral DNA by DNA polymerase gamma. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6206–6210. doi: 10.1073/pnas.78.10.6206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kollek R., Tseng B. Y., Goulian M. DNA polymerase requirements for parvovirus H-1 DNA replication in vitro. J Virol. 1982 Mar;41(3):982–989. doi: 10.1128/jvi.41.3.982-989.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kotin R. M., Linden R. M., Berns K. I. Characterization of a preferred site on human chromosome 19q for integration of adeno-associated virus DNA by non-homologous recombination. EMBO J. 1992 Dec;11(13):5071–5078. doi: 10.1002/j.1460-2075.1992.tb05614.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kotin R. M., Siniscalco M., Samulski R. J., Zhu X. D., Hunter L., Laughlin C. A., McLaughlin S., Muzyczka N., Rocchi M., Berns K. I. Site-specific integration by adeno-associated virus. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2211–2215. doi: 10.1073/pnas.87.6.2211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Laughlin C. A., Cardellichio C. B., Coon H. C. Latent infection of KB cells with adeno-associated virus type 2. J Virol. 1986 Nov;60(2):515–524. doi: 10.1128/jvi.60.2.515-524.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lebkowski J. S., McNally M. M., Okarma T. B., Lerch L. B. Adeno-associated virus: a vector system for efficient introduction and integration of DNA into a variety of mammalian cell types. Mol Cell Biol. 1988 Oct;8(10):3988–3996. doi: 10.1128/mcb.8.10.3988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. McLaughlin S. K., Collis P., Hermonat P. L., Muzyczka N. Adeno-associated virus general transduction vectors: analysis of proviral structures. J Virol. 1988 Jun;62(6):1963–1973. doi: 10.1128/jvi.62.6.1963-1973.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Miller A. D., Buttimore C. Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production. Mol Cell Biol. 1986 Aug;6(8):2895–2902. doi: 10.1128/mcb.6.8.2895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Miller A. D. Human gene therapy comes of age. Nature. 1992 Jun 11;357(6378):455–460. doi: 10.1038/357455a0. [DOI] [PubMed] [Google Scholar]
  17. Miller D. G., Adam M. A., Miller A. D. Gene transfer by retrovirus vectors occurs only in cells that are actively replicating at the time of infection. Mol Cell Biol. 1990 Aug;10(8):4239–4242. doi: 10.1128/mcb.10.8.4239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Miller D. G., Edwards R. H., Miller A. D. Cloning of the cellular receptor for amphotropic murine retroviruses reveals homology to that for gibbon ape leukemia virus. Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):78–82. doi: 10.1073/pnas.91.1.78. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mulligan R. C. The basic science of gene therapy. Science. 1993 May 14;260(5110):926–932. doi: 10.1126/science.8493530. [DOI] [PubMed] [Google Scholar]
  20. Muzyczka N. Use of adeno-associated virus as a general transduction vector for mammalian cells. Curr Top Microbiol Immunol. 1992;158:97–129. doi: 10.1007/978-3-642-75608-5_5. [DOI] [PubMed] [Google Scholar]
  21. Palmer T. D., Hock R. A., Osborne W. R., Miller A. D. Efficient retrovirus-mediated transfer and expression of a human adenosine deaminase gene in diploid skin fibroblasts from an adenosine deaminase-deficient human. Proc Natl Acad Sci U S A. 1987 Feb;84(4):1055–1059. doi: 10.1073/pnas.84.4.1055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pritchard C., Stout E. R., Bates R. C. Replication of parvoviral DNA. I. Characterization of a nuclear lysate system. J Virol. 1981 Jan;37(1):352–362. doi: 10.1128/jvi.37.1.352-362.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Robertson A. T., Stout E. R., Bates R. C. Aphidicolin inhibition of the production of replicative-form DNA during bovine parvovirus infection. J Virol. 1984 Mar;49(3):652–657. doi: 10.1128/jvi.49.3.652-657.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Roe T., Reynolds T. C., Yu G., Brown P. O. Integration of murine leukemia virus DNA depends on mitosis. EMBO J. 1993 May;12(5):2099–2108. doi: 10.1002/j.1460-2075.1993.tb05858.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ruffing M., Zentgraf H., Kleinschmidt J. A. Assembly of viruslike particles by recombinant structural proteins of adeno-associated virus type 2 in insect cells. J Virol. 1992 Dec;66(12):6922–6930. doi: 10.1128/jvi.66.12.6922-6930.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Samulski R. J. Adeno-associated virus: integration at a specific chromosomal locus. Curr Opin Genet Dev. 1993 Feb;3(1):74–80. doi: 10.1016/s0959-437x(05)80344-2. [DOI] [PubMed] [Google Scholar]
  27. Samulski R. J., Chang L. S., Shenk T. Helper-free stocks of recombinant adeno-associated viruses: normal integration does not require viral gene expression. J Virol. 1989 Sep;63(9):3822–3828. doi: 10.1128/jvi.63.9.3822-3828.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Samulski R. J., Zhu X., Xiao X., Brook J. D., Housman D. E., Epstein N., Hunter L. A. Targeted integration of adeno-associated virus (AAV) into human chromosome 19. EMBO J. 1991 Dec;10(12):3941–3950. doi: 10.1002/j.1460-2075.1991.tb04964.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Walsh C. E., Liu J. M., Xiao X., Young N. S., Nienhuis A. W., Samulski R. J. Regulated high level expression of a human gamma-globin gene introduced into erythroid cells by an adeno-associated virus vector. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):7257–7261. doi: 10.1073/pnas.89.15.7257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wolter S., Richards R., Armentrout R. W. Cell cycle-dependent replication of the DNA of minute virus of mice, a parvovirus. Biochim Biophys Acta. 1980 May 30;607(3):420–431. doi: 10.1016/0005-2787(80)90152-5. [DOI] [PubMed] [Google Scholar]
  31. Yakobson B., Koch T., Winocour E. Replication of adeno-associated virus in synchronized cells without the addition of a helper virus. J Virol. 1987 Apr;61(4):972–981. doi: 10.1128/jvi.61.4.972-981.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yalkinoglu A. O., Zentgraf H., Hübscher U. Origin of adeno-associated virus DNA replication is a target of carcinogen-inducible DNA amplification. J Virol. 1991 Jun;65(6):3175–3184. doi: 10.1128/jvi.65.6.3175-3184.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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