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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
. 1990 Mar;87(6):2211–2215. doi: 10.1073/pnas.87.6.2211

Site-specific integration by adeno-associated virus.

R M Kotin 1, M Siniscalco 1, R J Samulski 1, X D Zhu 1, L Hunter 1, C A Laughlin 1, S McLaughlin 1, N Muzyczka 1, M Rocchi 1, K I Berns 1
PMCID: PMC53656  PMID: 2156265

Abstract

Cellular sequences flanking integrated copies of the adeno-associated virus (AAV) genome were isolated from a latently infected clonal human cell line and used to probe genomic blots derived from an additional 21 independently derived clones of human cells latently infected with AAV. In genomic blots of uninfected human cell lines and of primary human tissue, each flanking-sequence probe hybridized to unique bands, but in 15 of the 22 latently infected clones the flanking sequences hybridized not only to the original fragments but also to a total of 36 additional species. AAV probes also hybridized to 22 of these new bands, representing 11 of the 15 positive clones, but never to the fragment characteristic of uninfected cell DNA. From these data we conclude that the AAV genome preferentially integrates into a specific region of the cellular genome. We have determined that the integration site is unique to chromosome 19 by somatic cell hybrid mapping, and this sequence has been isolated from uninfected human DNA.

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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. Casey G., Smith R., McGillivray D., Peters G., Dickson C. Characterization and chromosome assignment of the human homolog of int-2, a potential proto-oncogene. Mol Cell Biol. 1986 Feb;6(2):502–510. doi: 10.1128/mcb.6.2.502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cheung A. K., Hoggan M. D., Hauswirth W. W., Berns K. I. Integration of the adeno-associated virus genome into cellular DNA in latently infected human Detroit 6 cells. J Virol. 1980 Feb;33(2):739–748. doi: 10.1128/jvi.33.2.739-748.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  5. Hameister H., Ropers H. H., Grzeschik K. H. Assignment of the gene for human glucose dehydrogenase (E.C. 1.1.1.47) to chromosome 1 using somatic cell hybrids. Cytogenet Cell Genet. 1978;22(1-6):200–202. doi: 10.1159/000130935. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Hayward W. S., Neel B. G., Astrin S. M. Activation of a cellular onc gene by promoter insertion in ALV-induced lymphoid leukosis. Nature. 1981 Apr 9;290(5806):475–480. doi: 10.1038/290475a0. [DOI] [PubMed] [Google Scholar]
  8. Hermonat P. L., Labow M. A., Wright R., Berns K. I., Muzyczka N. Genetics of adeno-associated virus: isolation and preliminary characterization of adeno-associated virus type 2 mutants. J Virol. 1984 Aug;51(2):329–339. doi: 10.1128/jvi.51.2.329-339.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jessberger R., Heuss D., Doerfler W. Recombination in hamster cell nuclear extracts between adenovirus type 12 DNA and two hamster preinsertion sequences. EMBO J. 1989 Mar;8(3):869–878. doi: 10.1002/j.1460-2075.1989.tb03448.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kotin R. M., Berns K. I. Organization of adeno-associated virus DNA in latently infected Detroit 6 cells. Virology. 1989 Jun;170(2):460–467. doi: 10.1016/0042-6822(89)90437-6. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. Popescu N. C., Amsbaugh S. C., DiPaolo J. A. Human papillomavirus type 18 DNA is integrated at a single chromosome site in cervical carcinoma cell line SW756. J Virol. 1987 May;61(5):1682–1685. doi: 10.1128/jvi.61.5.1682-1685.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rocchi M., Roncuzzi L., Santamaria R., Archidiacono N., Dente L., Romeo G. Mapping through somatic cell hybrids and cDNA probes of protein C to chromosome 2, factor X to chromosome 13, and alpha 1-acid glycoprotein to chromosome 9. Hum Genet. 1986 Sep;74(1):30–33. doi: 10.1007/BF00278781. [DOI] [PubMed] [Google Scholar]
  15. Rose J. A., Berns K. I., Hoggan M. D., Koczot F. J. Evidence for a single-stranded adenovirus-associated virus genome: formation of a DNA density hybrid on release of viral DNA. Proc Natl Acad Sci U S A. 1969 Nov;64(3):863–869. doi: 10.1073/pnas.64.3.863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rose J. A., Koczot F. Adenovirus-associated virus multiplication. VII. Helper requirement for viral deoxyribonucleic acid and ribonucleic acid synthesis. J Virol. 1972 Jul;10(1):1–8. doi: 10.1128/jvi.10.1.1-8.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Shih C. C., Stoye J. P., Coffin J. M. Highly preferred targets for retrovirus integration. Cell. 1988 May 20;53(4):531–537. doi: 10.1016/0092-8674(88)90569-7. [DOI] [PubMed] [Google Scholar]
  18. Siegl G., Bates R. C., Berns K. I., Carter B. J., Kelly D. C., Kurstak E., Tattersall P. Characteristics and taxonomy of Parvoviridae. Intervirology. 1985;23(2):61–73. doi: 10.1159/000149587. [DOI] [PubMed] [Google Scholar]
  19. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]

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