Lentivirial Packaging Cell Lines

A series of lentiviral packaging cell lines for HIV based vectors are now available from ECACC. These have been developed by a group lead by Professor Mary Collins at University College London.

Researchers can introduce their gene of interest to be packaged into a human immunodeficiency virus type 1 (HIV-1) vector for production at high titres. HIV-1 vectors allow delivery of genes into a variety of cell types, both dividing and non-dividing. Previously it has been difficult to stably express large amounts of HIV-1 Gag-Pol and therefore no stable packaging cells were available to produce continuously high titre HIV-1 vectors. However, the cell lines displayed in Table 1 represent a stable HIV-1 packaging system.

These cell lines offer the opportunity to generate HIV vectors with a variety of different gammaretrovirus envelope proteins. The murine leukaemia virus amphotropic (MLA-A) envelope protein allows transduction of cells from a wide variety of species including human and mouse cells. The gibbon ape leukaemia virus (GALV) envelope protein is more efficient at transducing human haematopoetic cells and the feline endogenous virus RD114 envelope protein increases gene transfer efficiency into primary lymphocytes and CD34+ cells. Three 'producer' cell lines are available which produce disabled lentivirus carrying the green fluorescent protein (GFP) gene. These allow investigators to test the system - from virus production to transduction of target cells.

Table 1.  Lentiviral packaging cell lines for HIV based vectors.  GFP = Green Fluorescent Protein, GALV = gibbon ape leukaemia virus, RD114 = feline endogenous virus RD114, MLV-A = murine leukaemia virus amphotropic, Producer = cell line produces disabled lentivirus carrying the GFP gene and expressing the envelope protein noted, �synthetic Gag-Pol� is codon-optimised for efficient expression.

References:

Ikeda Y, Takeuchi Y, Martin F, Cosset FL, Mitrophanous K, Collins M: Continuous high-titer HIV-1 vector production. Nat Biotechnol 2003, 21(5):569-572.

Strang BL, Ikeda Y, Cosset FL, Collins MK, Takeuchi Y: Characterization of HIV-1 vectors with gammaretrovirus envelope glycoproteins produced from stable packaging cells. Gene Ther 2004, 11(7):591-598.