Russell Lab
Division of Hematology, School of Medicine, University of Washington
Foamy Reagents

 

 

 

 

 

Foamy virus (FV) vectors are a type of retroviral vector made from spumaviruses that can deliver a transgene cassette by integrating into the host cell genome. Wild-type FVs are not known to be pathogenic. There are several advantages of FV vectors, including a broad host range, large packaging capacity, stable pre-integration complex in non-dividing cells, a cDNA genome that is synthesized during vector production, resistance to inactivation by human complement, and efficient transduction of stem cells, including mesenchymal stem cells, hematopoietic stem cells, and embryonic stem cells. The deleted LTRs are silent in the vector backbone, so transgenes need to be expressed from an internal promoter. Unlike gammaretrovirus or lentivirus vectors, FV vectors do not preferentially integrate in genes. We have developed an FV vector production system that we are happy to distribute. This four-plasmid transient transfection system is described below. To obtain these plasmids please fill out and sign the Foamy Virus Vector Materials Transfer Agreement, and return it to our Technology Transfer Office as instructed on the form. We will then send you the reagents.

DF Vectors

DF vectors are "deleted foamy" virus vectors that only retain essential cis-acting viral sequences. The four plasmids requried to make DF vectors are pCiES, pCiGSDY, pCiPS and pDF. The first three plasmids are helper constructs expressing Env, Gag, and Pol respectively. pDF is a deleted foamy vector backbone with a polylinker for you to insert a transgene cassette. A description of the plasmids (Trobridge et al., 2002). Note however, that pCiGSDY os a newer version of the pCiGS Gag expression plasmid described in that paper, with a more complete deletion in the packaging signal (Josephson et al., 2004). All the plasmids encode ampicillin resistance.

For more information on foamy virus vectors see refernces below. See separate links for an FV Vector Production Protocol and the seuqences of plasmids.

pCiES,pCiGSDY, pCiPS and pDF (accurate as far as we know)

References

  Josephson , N.C. , Vassilopoulos, G., Trobridge, G.D., Priestley, G.V., Wood, B.L., Papayannopoulou, T., and Russell, D.W. (2002). Transduction of human NOD/SCID-repopulating cells with both lymphoid and myeloid potential by foamy virus vectors. Proc. Natl. Acad. Sci. USA 99:8295-8300.
  Josephson , N.C. , Trobridge, G., and Russell, D.W. (2004). Transduction of long-term and mobilized peripheral blood derived NOD/SCID repopulating cells by foamy virus vectors. Hum. Gene Ther. 15:87-92.
  Russell, D.W., and Miller, A.D. (1996). Foamy virus vectors. J. Virol. 70: 217-222.
  Trobridge, G., Josephson, N., Vassilopoulos, G., Mac, J, and Russell, D.W. (2002). Improved foamy virus vectors with minimal cis-acting sequences. Mol. Ther. 6:321-328.
  Trobridge, G.D., Vassilopoulos, G., Josephson, N., and Russell, D.W. (2002). Gene transfer with foamy virus vectors. Methods Enzymol. 346:628-648.
  Trobridge, G., and Russell, D.W. (2004). Cell cycle requirements for transduction by foamy virus vectors as compared to oncovirus and lentivirus vectors. J. Virol. 78:2327-2335.
  Trobridge, G.D., Miller, D.G., Jacobs, M.A., Allen, J.A., Kiem, H.P., Kaul, R., and Russell, D.W. (2006). Foamy virus vector integration sites in normal human cells. Proc. Natl. Acad. Sci. USA 103:1498-1503.
  Vassilopoulos, G., Trobridge, G., Josephson , N.C. , and Russell, D.W. (2001). Gene transfer into murine hematopoietic stem cells with helper-free foamy virus vectors. Blood 98:604-609.