AAV-Mediated Gene Delivery to the Lung

  • Laura P. van Lieshout
  • Jakob M. Domm
  • Sarah K. WoottonEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1950)


Adeno-associated virus (AAV) vectors are an efficient method of gene delivery to various tissues including the lung. Mouse models are often used as a preliminary preclinical model in order to advance AAV lung gene therapy vectors. In this chapter we describe an AAV purification protocol using heparin affinity chromatography as well as an intranasal and intratracheal method of delivering AAV vectors to the lungs of mice.

Key words

Adeno-associated virus AAV AAV6.2FF Lung gene therapy Lung delivery Lung gene transfer Heparin column chromatography AAV purification Intranasal instillation 



This work was supported by the Lung Association – Ontario and Cystic Fibrosis Canada.

Supplementary material

Video 1

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  1. 1.
    Zincarelli C, Soltys S, Rengo G, Rabinowitz J (2008) Analysis of AAV serotypes 1-9 mediated gene expression and tropism in mice after systemic injection. Mol Ther 16:1073–1080CrossRefGoogle Scholar
  2. 2.
    Zhong L, Li B, Mah CS, Govindasamy L, Agbandje-McKenna M, Coope M, Herzog RW, Zolotukhin I, Warrington KH, Weigel-Van Aken KA, Hobbs JA, Zolotukhin S, Muzyczka N, Srivastava A (2008) Next generation of adeno-associated virus 2 vectors: point mutations in tyrosines lead to high-efficiency transduction at lower doses. Proc Natl Acad Sci U S A 105:7827–7832CrossRefGoogle Scholar
  3. 3.
    Grimm D, Lee JS, Wang L, Desai T, Akache B, Storm TA, Kay MA (2008) In vitro and in vivo gene therapy vector evolution via multispecies interbreeding and retargeting of adeno-associated viruses. J Virol 82:5887–5911CrossRefGoogle Scholar
  4. 4.
    Li W, Zhang L, Wu Z, Pickles RJ, Samulski RJ (2011) AAV-6 mediated efficient transduction of mouse lower airways. Virology 417:327–333CrossRefGoogle Scholar
  5. 5.
    Halbert C, Madtes D, Vaughan A, Wang Z, Storb R, Tapscott SJ, Miller AD (2010) Expression of human alpha1-antitrypsin in mice and dogs following AAV6 vector-mediated gene transfer to the lungs. Mol Ther 18:1165–1172CrossRefGoogle Scholar
  6. 6.
    Halbert CL, Allen JM, Miller AD (2001) Adeno-associated virus type 6 (AAV6) vectors mediate efficient transduction of airway epithelial cells in mouse lungs compared to that of AAV2 vectors. J Virol 75:6615–6624CrossRefGoogle Scholar
  7. 7.
    Santry LA, Ingrao JC, Yu DL, de Jong JG, van Lieshout LP, Wood GA, Wootton SK (2017) AAV vector distribution in the mouse respiratory tract following four different methods of administration. BMC Biotechnol 17:43. Scholar
  8. 8.
    van Lieshout LP, Domm JM, Rindler TN, Frost KL, Sorenson DL, Medina SJ, Booth SA, Bridges JP, Wootton SK (2018) A novel triple-mutant AAV6 capsid induces rapid and potent transgene expression in the muscle and respiratory tract of mice. Mol Ther Methods Clin Dev 9:323–329CrossRefGoogle Scholar
  9. 9.
    van Lieshout LP, Soule G, Sorensen D, Frost KL, He S, Tierney K, Safronetz D, Booth SA, Kobinger GP, Qiu X, Wootton SK (2018) Intramuscular adeno-associated virus-mediated expression of monoclonal antibodies provides 100% protection against ebola virus infection in mice. J Infect Dis 217:916–925CrossRefGoogle Scholar
  10. 10.
    Aurnhammer C, Haase M, Muether N, Hausl M, Rauschhuber C, Huber I, Nitschko H, Busch U, Sing A, Ehrhardt A, Baiker A (2012) Universal real-time PCR for the detection and quantification of adeno-associated virus serotype 2-derived inverted terminal repeat sequences. Hum Gene Ther Methods 23:18–28CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Laura P. van Lieshout
    • 1
  • Jakob M. Domm
    • 1
  • Sarah K. Wootton
    • 1
    Email author
  1. 1.Department of PathobiologyUniversity of GuelphGuelphCanada

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