Adeno-Associated Virus

AAV is a single-stranded DNA parvovirus that commonly infects humans but has no known pathology. Recombinant adeno-associated virus has a wide host range, long-term expression in vivo, and low immunogenicity. Recombinant AAV is a promising viral vector for many gene transfer applications.

  • Tropism determined by choice of capsid serotype.
  • Long-term expression in terminally differentiated cells.
  • Wide range of cells can be infected, including non-dividing cells.
  • The size of the transgene cassette is limited to ~4.7 kb for non-self complementary vectors and ~2.2 kb for self-complementary vectors.

The University of Iowa VVC offers research grade, high quality AAV vectors with titers from 1 x1012 to 1 x1013 vg/ml. AAV vectors are prepared by either a dual baculovirus infection in SF-9 insect cells, or by a triple plasmid transfection in HEK-293 cells. The cis-acting AAV plasmid or baculovirus carries the AAV expression cassette containing the gene of interest flanked by the AAV inverted terminal repeats (ITR). The genes for the functional and structural proteins, rep and cap, respectively, and the adenovirus helper genes are provided in trans.

Purified vector preparations are subjected to a variety of quality control measures including a physical QPCR titer to asses viral genomes, transduced titer by FACS when reporters are available, and silver stain to further assess titer and viral purity. Control stocks are assayed for Cre recombinase and Luciferase activity.

The VVC currently offers the following serotypes: 1, 2, 5, 6, 8, 9, DJ, and rh10. Our nomenclature refers to the structural Rep proteins from AAV2 followed by the capsid serotype, for example AAV2/5. Novel peptide modified serotypes can also be produced on a case-by-case basis. Purification methods are customized for each serotype and may require optimization for novel capsids. Additional charges may apply for novel capsids.

Pilot studies may be needed to determine which serotype will work best in your system. We offer a serotype kit that includes 25 µl of AAV2/1, 2/2, 2/5, 2/6, 2/8, 2/9, 2/DJ, and 2/rh10 under the In-Stock Vector Page.

For more information on AAVDJ, please see:  Grimm D, Lee JS, Wang L, Desai T, Akache B, Storm TA, Kay MA. In vitro and in vivo gene therapy vector evolution via multispecies interbreeding and retargeting of adeno-associated viruses. J Virol. 2008;82:5887–5911.

For more information on AAVrh10, please see: 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 U S A. 200; 99(18):11854-9.