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Journal of Virology, June 1999, p. 4991-5000, Vol. 73, No. 6
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Minimum Requirements for Efficient Transduction of Dividing and
Nondividing Cells by Feline Immunodeficiency Virus Vectors
Julie C.
Johnston,1
Mehdi
Gasmi,1
Leland E.
Lim,2
John H.
Elder,3
Jiing-Kuan
Yee,1
Douglas J.
Jolly,1
Kevin P.
Campbell,2
Beverly L.
Davidson,4 and
Sybille L.
Sauter1,*
Center for Gene Therapy, Chiron Technologies,
San Diego, California 921211; Howard
Hughes Medical Institute and Departments of Physiology and Biophysics
and Neurology, University of Iowa College of
Medicine,2 and Department of
Internal Medicine, University of Iowa,4 Iowa
City, Iowa 52242; and Department of Molecular Biology, The
Scripps Research Institute, La Jolla, California
920373
Received 25 November 1998/Accepted 17 February 1999
The development of gene delivery vectors based on feline
immunodeficiency virus (FIV) is an attractive alternative to vectors based on primate sources for the delivery of genes into humans. To
investigate the requirements for efficient transduction of dividing and
nondividing cells by vector particles based on FIV, a series of
packaging and vector constructs was generated for which viral gene
expression was minimized and from which unnecessary cis-acting sequences were deleted. Pseudotyped vector
particles produced in 293T cells were used to transduce various target
cells, including contact-inhibited human skin fibroblasts and
growth-arrested HT1080 cells. FIV vectors in which the U3 promoter was
replaced with the cytomegalovirus promoter gave rise to over
50-fold-higher titers than FIV vectors containing the complete FIV 5'
long terminal repeat (LTR). Comparison of the transduction efficiencies
of vectors containing different portions of the FIV Gag coding region
indicates that at least a functional part of the FIV packaging signal
(
) is located within an area which includes the 5' LTR and the first 350 bp of gag. Transduction efficiencies of vectors
prepared without FIV vif and orf2 accessory
gene expression did not differ substantially from those of vectors
prepared with accessory gene expression in either dividing or
nondividing cells. The requirement for FIV rev-RRE was,
however, demonstrated by the inefficient production of vector particles
in the absence of rev expression. Together, these results
demonstrate the efficient transduction of nondividing cells in vitro by
a multiply attenuated FIV vector and contribute to an understanding of
the minimum requirements for efficient vector production and
infectivity. In addition, we describe the ability of an FIV vector to
deliver genes in vivo into hamster muscle tissue.
*
Corresponding author. Mailing address: Chiron
Technologies, Center for Gene Therapy, 11055 Roselle St., San
Diego, CA 92121. Phone: (619) 452-1288. Fax: (619) 623-9975. E-mail:
sybille_sauter{at}cc.chiron.com.
Journal of Virology, June 1999, p. 4991-5000, Vol. 73, No. 6
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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