Russell Lab
Division of Hematology, School of Medicine, University of Washington
AAV Serotype Reagents

 

 

 

 

 

There are several different serotypes of Adeno-Associated Virus (AAV).  Type 2 is the original and best-studied serotype. Our laboratory has generated infectious clones of two other serotypes: 3B and 6 (Rutledge et al., 1998).  Vectors based on AAV6 have several advantages over conventional AAV2 vectors, including excellent transduction of muscle, lung, liver and other tissues in vivo (Halbert et al., 2000).  AAV3B is especially useful for transducing pluripotent stem cells (Mitsui et al., 2009). AAV vectors can be packaged in alternative capsids by expressing the AAV6 or AAV3B cap genes during vector production.  We have generated 3 plasmids that can be used for this purpose. pRepCap6 contains both rep and cap genes from the AAV6 genome, but lacks the viral terminal repeats (Rutledge et al., 1998). It lacks adenovirus helper functions required for stock production. pDGM6 contains the AAV6 cap genes, AAV2 rep genes, and also adenovirus helper genes  (Gregorevic et al., 2004). pDGM3B contains the AAV3B cap genes, AAV2 rep genes, and also adenovirus helper genes (Khan et al., 2011).  To obtain any of these plasmids please fill out and submit the online Materials Transfer Agreement that can be found at the following UW MTA link, using the exact name for the materials as found in our publications. Once you have obtained approval from our Center for Commercialization, we will then send you the reagents.  See specific links for the sequences of pRepCap6, pDGM6, and pDGM3B. Please note that in the case of pDGM6 and pDGM3B, these are compiled sequences that may contain errors. We use the following diagnostic enzymes to check pDGM6: AatII (10070, 3533, 3327, 2967, and 1945 bp fragments seen) and MfeI (9244, 7695 and 4903 bp fragments). We have used the following diagnostic enzymes to check pDGM3B: AatII (10070, 4412, 3327, 2097, and 1945 bp fragments seen) and MfeI (16,948 and 4903 bp fragments). The pRepCap6 sequence is accurate as far as we know. Both plasmids confer ampicillin resistance.

References

  Gregorevic, P., Blankenship, M.J., Allen, J. M., Crawford, R.W., Meuse , L., Miller, D.G., Russell, D.W., and Chamberlain, J.S. (2004). Systemic Delivery of genes to striated muscles using adeno-associated virus vectors. Nat. Med. 10, 828-834.
  Halbert, C. L., Allen, J. M., and Miller, A. D. (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, 6615-6624.
  Khan, I.F., Hirata, R.K., and Russell, D.W. (2011). AAV-mediated gene targeting methods for human cells. Nature protocols 6, 482-501.
  Mitsui, K., Suzuki, K., Aizawa, E., Kawase, E., Suemori, H., Nakatsuji, N., and Mitani, K. (2009). Gene targeting in human pluripotent stem cells with adeno-associated virus vectors. Biochem Biophys Res Commun 388, 711-717.
  Rutledge, E. A., Halbert, C. L., and Russell, D. W. (1998). Infectious clones and vectors derived from adeno-associated virus (AAV) serotypes other than AAV type 2. J Virol 72, 309-319.