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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 (Psi ) 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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