An all-feline retroviral packaging system for transduction of human cells.
Abstract The subgroup C feline leukemia virus (FeLV-C) receptor FLVCR is a widely expressed 12-transmembrane domain transporter that exports cytoplasmic heme and is a promising target for retrovirus-mediated gene delivery. Previous studies demonstrated that FeLV-C pseudotype vectors were more efficient at targeting human hematopoietic stem cells than those pseudotyped with gibbon ape leukemia virus (GALV), and thus we developed an all FeLV-C-based packaging system, termed CatPac. CatPac is helper-virus free and can produce higher titer vectors than existing gammaretroviral packaging systems, including systems mixing Moloney murine leukemia virus (MoMLV) Gag-Pol and FeLV-C Env proteins. The vectors can be readily concentrated (>30-fold), refrozen (three to five times), and held on ice (>2 days) with little loss of titer. Furthermore, we demonstrate that CatPac pseudotype vectors efficiently target early CD34(+)CD38(-) stem/progenitor cells, monocytic and erythroid progenitors, activated T cells, mature macrophages, and cancer cell lines, suggesting utility for human cell and cell line transduction and possibly gene therapy.
Doty
Raymond T
RT
Department of Medicine, University of Washington, Seattle, WA 98195, USA.
Sabo
Kathleen M
KM
Chen
Jing
J
Miller
A Dusty
AD
Abkowitz
Janis L
JL
D000818:Animals; D018952:Antigens, CD34 / Q000235:genetics / Q000378:metabolism; D002415:Cats; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells / Q000378:metabolism; D006801:Humans; D005256:Leukemia Virus, Feline / Q000235:genetics*; D026901:Membrane Transport Proteins / Q000235:genetics*; D011991:Receptors, Virus / Q000235:genetics*; D014161:Transduction, Genetic
D018952:Antigens, CD34; C492690:FLVCR1 protein, human; D026901:Membrane Transport Proteins; D011991:Receptors, Virus