Efficient transduction of nondividing cells by optimized feline immunodeficiency virus vectors.
Second- and third-generation three-plasmid vector systems, termed FELIX, were constructed from feline immunodeficiency virus (FIV). To enhance vector production, the weak FIV long terminal repeat promoter was replaced with the human cytomegalovirus enhancer/promoter. To construct a minimal system in which Gag-Pol was the only viral protein present, the cytoplasmic transport element was used in place of the FIV Rev-RRE system to facilitate nuclear export of Gag-Pol and the transfer vector. Unconcentrated vector titers routinely exceeded 1 x 10(6) IU/mL for most constructs tested. Second- and optimized third-generation vectors were capable of efficiently infecting G1/S- and G2/M-arrested cells. FIV-based FELIX vectors transduced human dendritic cells, hepatocytes, and aortic smooth muscle with efficiencies similar to that of a control 3T3 cell line. All three of these primary cell types were transducible by both the second- and third-generation FELIX vectors, demonstrating that FIV Gag-Pol alone contains the determinants necessary for transduction of primary cells. In cross-packaging tests, we observed that HIV Gag-Pol does not substantially package FIV vectors; consequently, use of such vectors in human immunodeficiency virus-infected cells should not lead to efficient mobilization of the inserted gene. Thus, this FIV-based vector system offers high efficiency and stable delivery of genes to numerous nondividing and primary cell types, opening new avenues for biological inquiry into normal human cells.
Program in Immunology, Stanford University Medical Center, California 94305-5332, USA. curran@leland.stanford.edu
M A
MA
Curran
S M
SM
Kaiser
P L
PL
Achacoso
G P
GP
Nolan
D016475:3T3 Cells; D000818:Animals; D002415:Cats; D002460:Cell Line; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics; D004742:Enhancer Elements, Genetic; D015685:Fusion Proteins, gag-pol / Q000235:genetics; D015316:Genetic Therapy; D005822:Genetic Vectors; D006678:HIV / Q000235:genetics; D006801:Humans; D016089:Immunodeficiency Virus, Feline / Q000235:genetics*; D007399:Interphase; D009052:Leukemia Virus, Murine / Q000235:genetics; D051379:Mice; D011401:Promoter Regions, Genetic; D020079:Terminal Repeat Sequences; D014161:Transduction, Genetic
D015685:Fusion Proteins, gag-pol