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.
Curran
M A
MA
Program in Immunology, Stanford University Medical Center, California 94305-5332, USA. curran@leland.stanford.edu
Kaiser
S M
SM
Achacoso
P L
PL
Nolan
G P
GP
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