Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Short Communication
  • Published:

Assessment of ocular transduction using single-stranded and self-complementary recombinant adeno-associated virus serotype 2/8

Gene Therapy volume 15, pages 463–467 (2008)Cite this article

Abstract

To date adeno-associated viral (AAV) vectors are the only gene therapy vectors that have been shown to efficiently transduce photoreceptor cells and have thus become the most commonly used vector for ocular transduction. Various AAV serotypes have been evaluated in the eye, the first of which was AAV2, which is able to transduce photoreceptors, retinal pigment epithelium (RPE) and retinal ganglion cells. AAV serotypes 1 and 4, as well as AAV2 pseudotyped with these capsids, only transduce the RPE. AAV serotype 5 and AAV2/5 transduce the photoreceptors as well as RPE, but not retinal ganglion cells. Here, we assessed the capacity of the novel serotype AAV2/8 to transduce various ocular tissues of the adult murine retina by administering AAV2/8 green fluorescent protein intravitreally, subretinally and intracamerally. We also determined the kinetics and efficiency of self-complementary AAV (scAAV) vectors of serotypes 2/2, 2/5 and 2/8 and compared them with single-stranded AAV (ssAAV). We found that ssAAV2/8 transduces photoreceptors and RPE more efficiently than ssAAV2/2 and ssAAV2/5, and that scAAV2/8 had faster onset and higher transgene expression than ssAAV2/8. This improved transduction efficiency might facilitate the development of improved gene therapy protocols for inherited retinal degenerations, particularly those caused by defects in photoreceptor-specific genes.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4

Similar content being viewed by others

References

  1. Ali RR . Prospects for gene therapy. Novartis Found Symp 2004; 255: 165–172.

    CAS  PubMed  Google Scholar 

  2. Bainbridge JW, Tan MH, Ali RR . Gene therapy progress and prospects: the eye. Gene Therapy 2006; 13: 1191–1197.

    Article  CAS  Google Scholar 

  3. Goncalves MA . Adeno-associated virus: from defective virus to effective vector. Virol J 2005; 2: 43–44.

    Article  Google Scholar 

  4. Sarra GM, Stephens C, Schlichtenbrede FC, Bainbridge JW, Thrasher AJ, Luthert PJ et al. Kinetics of transgene expression in mouse retina following sub-retinal injection of recombinant adeno-associated virus. Vision Res 2002; 42: 541–549.

    Article  CAS  Google Scholar 

  5. Yang GS, Schmidt M, Yan Z, Lindbloom JD, Harding TC, Donahue BA et al. Virus-mediated transduction of murine retina with adeno-associated virus: effects of viral capsid and genome size. J Virol 2002; 76: 7651–7660.

    Article  CAS  Google Scholar 

  6. Auricchio A, Kobinger G, Anand V, Hildinger M, O'Connor E, Maguire AM et al. Exchange of surface proteins impacts on viral vector cellular specificity and transduction characteristics: the retina as a model. Hum Mol Genet 2001; 10: 3075–3081.

    Article  CAS  Google Scholar 

  7. Auricchio A, Hildinger M, O'Connor E, Gao GP, Wilson JM . Isolation of highly infectious and pure adeno-associated virus type 2 vectors with a single-step gravity-flow column. Hum Gene Ther 2001; 12: 71–76.

    Article  CAS  Google Scholar 

  8. Auricchio A . Pseudotyped AAV vectors for constitutive and regulated gene expression in the eye. Vision Res 2003; 43: 913–918.

    Article  CAS  Google Scholar 

  9. Weber M, Rabinowitz J, Provost N, Conrath H, Folliot S, Briot D et al. Recombinant adeno-associated virus serotype 4 mediates unique and exclusive long-term transduction of retinal pigmented epithelium in rat, dog, and nonhuman primate after subretinal delivery. Mol Ther 2003; 7: 774–781.

    Article  CAS  Google Scholar 

  10. Ding W, Zhang L, Yan Z, Engelhardt JF . Intracellular trafficking of adeno-associated viral vectors. Gene Therapy 2005; 12: 873–880.

    Article  CAS  Google Scholar 

  11. Thomas CE, Storm TA, Huang Z, Kay MA . Rapid uncoating of vector genomes is the key to efficient liver transduction with pseudotyped adeno-associated virus vectors. J Virol 2004; 78: 3110–3122.

    Article  CAS  Google Scholar 

  12. Vincent-Lacaze N, Snyder RO, Gluzman R, Bohl D, Lagarde C, Danos O . Structure of adeno-associated virus vector DNA following transduction of the skeletal muscle. J Virol 1999; 73: 1949–1955.

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Wang Z, Ma HI, Li J, Sun L, Zhang J, Xiao X . Rapid and highly efficient transduction by double-stranded adeno-associated virus vectors in vitro and in vivo. Gene Therapy 2003; 10: 2105–2111.

    Article  CAS  Google Scholar 

  14. Broekman ML, Comer LA, Hyman BT, Sena-Esteves M . Adeno-associated virus vectors serotyped with AAV8 capsid are more efficient than AAV-1 or -2 serotypes for widespread gene delivery to the neonatal mouse brain. Neuroscience 2006; 138: 501–510.

    Article  CAS  Google Scholar 

  15. Inagaki K, Fuess S, Storm TA, Gibson GA, Mctiernan CF, Kay MA et al. Robust systemic transduction with AAV9 vectors in mice: efficient global cardiac gene transfer superior to that of AAV8. Mol Ther 2006; 14: 45–53.

    Article  CAS  Google Scholar 

  16. Wang Z, Zhu T, Rehman KK, Bertera S, Zhang J, Chen C et al. Widespread and stable pancreatic gene transfer by adeno-associated virus vectors via different routes. Diabetes 2006; 55: 875–884.

    Article  CAS  Google Scholar 

  17. Davidoff AM, Gray JT, Ng CY, Zhang Y, Zhou J, Spence Y et al. Comparison of the ability of adeno-associated viral vectors pseudotyped with serotype 2, 5, and 8 capsid proteins to mediate efficient transduction of the liver in murine and nonhuman primate models. Mol Ther 2005; 11: 875–888.

    Article  CAS  Google Scholar 

  18. Nathwani AC, Gray JT, Ng CY, Zhou J, Spence Y, Waddington SN et al. Self-complementary adeno-associated virus vectors containing a novel liver-specific human factor IX expression cassette enable highly efficient transduction of murine and nonhuman primate liver. Blood 2006; 107: 2653–2661.

    Article  CAS  Google Scholar 

  19. Ali RR, Sarra GM, Stephens C, Alwis MD, Bainbridge JW, Munro PM et al. Restoration of photoreceptor ultrastructure and function in retinal degeneration slow mice by gene therapy. Nat Genet 2000; 25: 306–310.

    Article  CAS  Google Scholar 

  20. Zhang X, De Alwis M, Hart SL, Fitzke FW, Inglis SC, Boursnell ME et al. High-titer recombinant adeno-associated virus production from replicating amplicons and herpes vectors deleted for glycoprotein H. Hum Gene Ther 1999; 10: 2527–2537.

    Article  CAS  Google Scholar 

  21. Clark KR, Liu X, McGrath JP, Johnson PR . Highly purified recombinant adeno-associated virus vectors are biologically active and free of detectable helper and wild-type viruses. Hum Gene Ther 1999; 10: 1031–1039.

    Article  CAS  Google Scholar 

  22. Davidoff AM, Ng CY, Sleep S, Gray J, Azam S, Zhao Y et al. Purification of recombinant adeno-associated virus type 8 vectors by ion exchange chromatography generates clinical grade vector stock. J Virol Methods 2004; 121: 209–215.

    Article  CAS  Google Scholar 

  23. Akache B, Grimm D, Shen X, Fuess S, Yant SR, Glazer DS et al. A two-hybrid screen identifies cathepsins B and L as uncoating factors for adeno-associated virus 2 and 8. Mol Ther 2007; 15: 330–339.

    Article  CAS  Google Scholar 

  24. Gao GP, Alvira MR, Wang L, Calcedo R, Johnston J, Wilson JM . Novel adeno-associated viruses from rhesus monkeys as vectors for human gene therapy. Proc Natl Acad Sci USA 2002; 99: 11854–11859.

    Article  CAS  Google Scholar 

  25. Wang L, Cao O, Swalm B, Dobrzynski E, Mingozzi F, Herzog RW . Major role of local immune responses in antibody formation to factor IX in AAV gene transfer. Gene Therapy 2005; 12: 1453–1464.

    Article  CAS  Google Scholar 

  26. Allocca M, Mussolino C, Hoyos MG, Sanges D, Iodice C, Petrillo M et al. Novel AAV serotypes efficiently transduce murine photoreceptors. J Virol 2007; 81: 11372–11380.

    Article  CAS  Google Scholar 

  27. Ren C, Kumar S, Shaw DR, Ponnazhagan S . Genomic stability of self-complementary adeno-associated virus 2 during early stages of transduction in mouse muscle in vivo. Hum Gene Ther 2005; 16: 1047–1057.

    Article  CAS  Google Scholar 

  28. Buch PK, MacLaren RE, Duran Y, Balaggan KS, MacNeil A, Schlichtenbrede FC et al. In contrast to AAV-mediated Cntf expression, AAV-mediated Gdnf expression enhances gene replacement therapy in rodent models of retinal degeneration. Mol Ther 2006; 14: 700–709.

    Article  CAS  Google Scholar 

  29. Broderick CA, Smith AJ, Balaggan KS, Georgarias A, Buch PK, Trittibach PC et al. Local administration of an adeno-associated viral vector expressing IL-10 reduces monocyte infiltration and subsequent photoreceptor damage during experimental autoimmune uveitis. Mol Ther 2005; 12: 369–373.

    Article  CAS  Google Scholar 

  30. Ali RR, Reichel MB, Thrasher AJ, Levinsky RJ, Kinnon C, Kanuga N et al. Gene transfer into the mouse retina mediated by an adeno-associated viral vector. Hum Mol Genet 1996; 5: 591–594.

    Article  CAS  Google Scholar 

  31. Bennett J, Duan D, Engelhardt JF, Maguire AM . Real-time, noninvasive in vivo assessment of adeno-associated virus-mediated retinal transduction. Invest Ophthalmol Vis Sci 1997; 38: 2857–2863.

    CAS  PubMed  Google Scholar 

  32. Grant CA, Ponnazhagan S, Wang XS, Srivastava A, Li T . Evaluation of recombinant adeno-associated virus as a gene transfer vector for the retina. Curr Eye Res 1997; 16: 949–956.

    Article  CAS  Google Scholar 

  33. Lai YK, Rolling F, Baker E, Rakoczy PE . Kinetics of efficient recombinant adeno-associated virus transduction in retinal pigment epithelial cells. Exp Cell Res 2001; 267: 184–192.

    Article  CAS  Google Scholar 

  34. Rolling F, Shen WY, Tabarias H, Constable I, Kanagasingam Y, Barry CJ et al. Evaluation of adeno-associated virus-mediated gene transfer into the rat retina by clinical fluorescence photography. Hum Gene Ther 1999; 10: 641–648.

    Article  CAS  Google Scholar 

  35. Dudus L, Anand V, Acland GM, Chen SJ, Wilson JM, Fisher KJ et al. Persistent transgene product in retina, optic nerve and brain after intraocular injection of rAAV. Vision Res 1999; 39: 2545–2553.

    Article  CAS  Google Scholar 

  36. Smith AJ, Schlichtenbrede FC, Tschernutter M, Bainbridge JW, Thrasher AJ, Ali RR . AAV-mediated gene transfer slows photoreceptor loss in the RCS rat model of retinitis pigmentosa. Mol Ther 2003; 8: 188–195.

    Article  CAS  Google Scholar 

  37. Acland GM, Aguirre GD, Ray J, Zhang Q, Aleman TS, Cideciyan AV et al. Gene therapy restores vision in a canine model of childhood blindness. Nat Genet 2001; 28: 92–95.

    CAS  PubMed  Google Scholar 

  38. Schlichtenbrede FC, Da Cruz L, Stephens C, Smith AJ, Georgiadis A, Thrasher AJ et al. Long-term evaluation of retinal function in Prph2Rd2/Rd2 mice following AAV-mediated gene replacement therapy. J Gene Med 2003; 5: 757–764.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the European Commission (EVI-Genoret and Clinigene) and a grant from the department of health and from the Special Trustees of Moorfields Eye Hospital to RRA and a grant from the Katharine Dormandy Trust, UK to ACN.

Author information

Author notes

  1. M Natkunarajah and P Trittibach: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Molecular Therapy, UCL Institute of Ophthalmology, University College London, London, UK

    M Natkunarajah, Y Duran, S E Barker, A J Smith & R R Ali

  2. Department of Ophthalmology, University of Bern, Inselspital, Bern, Switzerland

    P Trittibach

  3. Department of Haematology, University College London, London, UK

    J McIntosh & A C Nathwani

Authors
  1. M Natkunarajah
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. P Trittibach
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J McIntosh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Y Duran
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. S E Barker
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. A J Smith
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. A C Nathwani
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. R R Ali
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to A J Smith.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Natkunarajah, M., Trittibach, P., McIntosh, J. et al. Assessment of ocular transduction using single-stranded and self-complementary recombinant adeno-associated virus serotype 2/8. Gene Ther 15, 463–467 (2008). https://doi.org/10.1038/sj.gt.3303074

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3303074

Keywords

This article is cited by

Search

Advanced search

Quick links