Adeno-Associated Virus (AAV)

Background

Adeno-Associated Virus (AAV) is a nonpathogenic virus species that belongs to the Parvoviridae family. AAV is classified as small (25nm) and contains a single-stranded nonenveloped DNA genome. Infection with AAV occurs only with the help of other viruses, either herpesvirus or adenovirus (hence adeno-associated virus), causing only a very mild immune response in humans. There are twelve serotypes of human AAV but the number of nonhuman AAVs exceeds 100. AAV2 is the only mammalian DNA virus that is known to integrate in a specific site of the genome.

AAV in Gene Therapy

The low immunogenicity and the ability to infect dividing as well as non-dividing cells with stable expression make the adeno-associated virus an attractive vector for the application in gene therapy. AAV has successfully been proven as gene therapy vector with the ability to attach and enter the target cell, transfer to the nucleus and express the transgene in a stable manner over a sustained period of time. In several clinical trials (e.g. FIX, CFTR, Parkinsons’s, Canavan disease) AAV did not show any serious vector-related adverse effects. Clinically relevant tissues have, however, revealed a diversified susceptibility to AAV infection. The gene transduction with AAV2 vector in muscle, retina, liver and heart resulted in lower gene expression compared to AAV serotypes 1, 5, 8 and 9 transduction in the respective tissues. The challenge of using AAV as gene therapy vector, however, lies in the resistance of some tissues to transduction with the available AAV serotypes.

Characterization of AAV preparations

With the increasing importance of AAV vectors in the field of gene therapy there is an increasing demand for robust & reliable methods for rAAV characterization. At this moment, quantification methods for rAAV vector preparations currently include quantitative PCR (qPCR) & digital droplet PCR (ddPCR) for the determination of the DNA titer as well as dot blot &titration ELISAfor the determination of the total capsid titer (assembled viral capsids proteins). Methods using electron microscopy, protein chromatography, flow cytometry, or virus particle counting instruments have also been described, but are generally only applicable for highly skilled users and/or require specialised and expensive equipment. Find out moreabout currently used methods and a comparison of AAV quantification methods. 

You may enrich your research on AAV by browsing through PROGEN's AAV Product portfolio and access the highest quality research tools.

For more information about PROGEN´s antibodies and AAV titration ELISAs, see PROGEN´s posters in the poster download area below.

Target-Organs for AAV

  1. Xiao, W. et al. Gene therapy vectors based on adeno-associated virus type 1. J. Virol.73, 3994–4003 (1999).
  2. Manfredsson, F. P., Rising, A. C. & Mandel, R. J. AAV9: A potential blood-brain barrier buster. Molecular Therapy17, 403–405 (2009).
  3. Sen, D. et al. Improved adeno-associated virus (AAV) serotype 1 and 5 vectors for gene therapy. Sci. Rep.6, (2013).
  4. Zabner, J. et al. Adeno-associated virus type 5 (AAV5) but not AAV2 binds to the apical surfaces of airway epithelia and facilitates gene transfer. J. Virol.74, 3852–3858 (2000).
  5. Shen, S., Troupes, A. N., Pulicherla, N. & Asokan, A. Multiple Roles for Sialylated Glycans in Determining the Cardiopulmonary Tropism of Adeno-Associated Virus 4. J. Virol.87, 13206–13213 (2013).
  6. Ai, J. et al. Adeno-associated virus serotype rh.10 displays strong muscle tropism following intraperitoneal delivery. Sci. Rep.7, (2017).
  7. Cheng, B. et al. Development of optimized AAV3 serotype vectors: Mechanism of high-efficiency transduction of human liver cancer cells. Gene Ther.19, 375–384 (2012).
  8. Mingozzi, F. & High, K. A. Immune responses to AAV vectors: Overcoming barriers to successful gene therapy. Blood122, 23–36 (2013).
  9. Richter, M. et al. Adeno-associated virus vector transduction of vascular smooth muscle cells in vivo. Physiol. Genomics2, 117–27 (2000).
  10. Mori, S. et al. Tissue distribution of cynomolgus adeno-associated viruses AAV10, AAV11, and AAVcy.7 in naturally infected monkeys. Arch. Virol.153, 375–380 (2008).
  11. Guo, Y. et al. A Single Injection of Recombinant Adeno-Associated Virus into the Lumbar Cistern Delivers Transgene Expression Throughout the Whole Spinal Cord. Mol. Neurobiol. (2016).
  12. Schmidt, M. et al. Adeno-Associated Virus Type 12 (AAV12): a Novel AAV Serotype with Sialic Acid- and Heparan Sulfate Proteoglycan-Independent Transduction Activity. J. Virol.82, 1399–1406 (2008).

Poster Downloads

Check out PROGEN´s posters about the development of reliable AAV standards, the new recombinant AAV2 antibody and comparison of negative staining EM and cryo-EM for the determination of the ratio of full and empty AAV capsids.