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Adeno-Associated Virus as Gene Delivery Vehicle into the Retina

  • Shuyun Deng
  • Kazuhiro OkaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2092)

Abstract

Initially discovered as a contaminant of adenovirus preparations, adeno-associated virus (AAV) has proved one of the most promising viral vectors for human gene therapy. The safety profile of AAV has been well-characterized in vivo studies, and the first gene therapy for patients with vision loss caused by Leber congenital amaurosis or retinitis pigmentosa was approved by the US Food and Drug Administration in 2017. This is an exciting era for investigators working on retina biology and treatments for blindness. In this chapter, we provide detailed methods for laboratory-scale production, purification, and characterization of AAV.

Key words

Adeno-associated virus Serotype Packaging Purification Titration 

Notes

Acknowledgment

We thank Dr. Sean Hartig for helpful discussion and critical reading of the manuscript. This work was supported by Gene Vector Core, Baylor College of Medicine Advanced Technology Cores.

References

  1. 1.
    Atchison RW, Casto BC, Hammon WM (1965) Adenovirus-associated defective virus particles. Science 149(3685):754–756CrossRefGoogle Scholar
  2. 2.
    Pillay S, Carette JE (2017) Host determinants of adeno-associated viral vector entry. Curr Opin Virol 24:124–131.  https://doi.org/10.1016/j.coviro.2017.06.003CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Dalkara D, Byrne LC, Klimczak RR, Visel M, Yin L, Merigan WH, Flannery JG, Schaffer DV (2013) In vivo-directed evolution of a new adeno-associated virus for therapeutic outer retinal gene delivery from the vitreous. Sci Transl Med 5(189):189ra176.  https://doi.org/10.1126/scitranslmed.3005708CrossRefGoogle Scholar
  4. 4.
    Ramachandran PS, Lee V, Wei Z, Song JY, Casal G, Cronin T, Willett K, Huckfeldt R, Morgan JI, Aleman TS, Maguire AM, Bennett J (2017) Evaluation of dose and safety of AAV7m8 and AAV8BP2 in the non-human primate retina. Hum Gene Ther 28(2):154–167.  https://doi.org/10.1089/hum.2016.111CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Grimm D, Zolotukhin S (2015) E pluribus Unum: 50 years of research, millions of viruses, and one goal—tailored acceleration of AAV evolution. Mol Ther 23(12):1819–1831.  https://doi.org/10.1038/mt.2015.173CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Weinmann J, Grimm D (2017) Next-generation AAV vectors for clinical use: an ever-accelerating race. Virus Genes 53(5):707–713.  https://doi.org/10.1007/s11262-017-1502-7CrossRefPubMedGoogle Scholar
  7. 7.
    Colella P, Ronzitti G, Mingozzi F (2018) Emerging issues in AAV-mediated in vivo gene therapy. Mol Ther Methods Clin Dev 8:87–104.  https://doi.org/10.1016/j.omtm.2017.11.007CrossRefPubMedGoogle Scholar
  8. 8.
    Nakai H, Yant SR, Storm TA, Fuess S, Meuse L, Kay MA (2001) Extrachromosomal recombinant adeno-associated virus vector genomes are primarily responsible for stable liver transduction in vivo. J Virol 75(15):6969–6976.  https://doi.org/10.1128/JVI.75.15.6969-6976.2001CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Bell P, Moscioni AD, McCarter RJ, Wu D, Gao G, Hoang A, Sanmiguel JC, Sun X, Wivel NA, Raper SE, Furth EE, Batshaw ML, Wilson JM (2006) Analysis of tumors arising in male B6C3F1 mice with and without AAV vector delivery to liver. Mol Ther 14(1):34–44.  https://doi.org/10.1016/j.ymthe.2006.03.008CrossRefPubMedGoogle Scholar
  10. 10.
    Zhong L, Malani N, Li M, Brady T, Xie J, Bell P, Li S, Jones H, Wilson JM, Flotte TR, Bushman FD, Gao G (2013) Recombinant adeno-associated virus integration sites in murine liver after ornithine transcarbamylase gene correction. Hum Gene Ther 24(5):520–525.  https://doi.org/10.1089/hum.2012.112CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Nault JC, Datta S, Imbeaud S, Franconi A, Mallet M, Couchy G, Letouze E, Pilati C, Verret B, Blanc JF, Balabaud C, Calderaro J, Laurent A, Letexier M, Bioulac-Sage P, Calvo F, Zucman-Rossi J (2015) Recurrent AAV2-related insertional mutagenesis in human hepatocellular carcinomas. Nat Genet 47(10):1187–1193.  https://doi.org/10.1038/ng.3389CrossRefPubMedGoogle Scholar
  12. 12.
    Dong B, Nakai H, Xiao W (2010) Characterization of genome integrity for oversized recombinant AAV vector. Mol Ther 18(1):87–92.  https://doi.org/10.1038/mt.2009.258CrossRefPubMedGoogle Scholar
  13. 13.
    Ghosh A, Duan D (2007) Expanding adeno-associated viral vector capacity: a tale of two vectors. Biotechnol Genet Eng Rev 24:165–177CrossRefGoogle Scholar
  14. 14.
    Maddalena A, Tornabene P, Tiberi P, Minopoli R, Manfredi A, Mutarelli M, Rossi S, Simonelli F, Naggert JK, Cacchiarelli D, Auricchio A (2018) Triple vectors expand AAV transfer capacity in the retina. Mol Ther 26(2):524–541.  https://doi.org/10.1016/j.ymthe.2017.11.019CrossRefPubMedGoogle Scholar
  15. 15.
    Ayuso E, Mingozzi F, Montane J, Leon X, Anguela XM, Haurigot V, Edmonson SA, Africa L, Zhou S, High KA, Bosch F, Wright JF (2010) High AAV vector purity results in serotype- and tissue-independent enhancement of transduction efficiency. Gene Ther 17(4):503–510.  https://doi.org/10.1038/gt.2009.157CrossRefPubMedGoogle Scholar
  16. 16.
    Lock M, Alvira M, Vandenberghe LH, Samanta A, Toelen J, Debyser Z, Wilson JM (2010) Rapid, simple, and versatile manufacturing of recombinant adeno-associated viral vectors at scale. Hum Gene Ther 21(10):1259–1271.  https://doi.org/10.1089/hum.2010.055CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Wright JF, Le T, Prado J, Bahr-Davidson J, Smith PH, Zhen Z, Sommer JM, Pierce GF, Qu G (2005) Identification of factors that contribute to recombinant AAV2 particle aggregation and methods to prevent its occurrence during vector purification and formulation. Mol Ther 12(1):171–178.  https://doi.org/10.1016/j.ymthe.2005.02.021CrossRefPubMedGoogle Scholar
  18. 18.
    D’Costa S, Blouin V, Broucque F, Penaud-Budloo M, Francois A, Perez IC, Le Bec C, Moullier P, Snyder RO, Ayuso E (2016) Practical utilization of recombinant AAV vector reference standards: focus on vector genomes titration by free ITR qPCR. Mol Ther Methods Clin Dev 5:16019.  https://doi.org/10.1038/mtm.2016.19CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Advanced Technology CoresBaylor College of MedicineHoustonUSA
  2. 2.Department of Molecular and Cellular BiologyBaylor College of MedicineHoustonUSA

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