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Journal of Virology, October 2001, p. 9339-9344, Vol. 75, No. 19
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.19.9339-9344.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Development of an Avian Leukosis-Sarcoma Virus Subgroup A Pseudotyped Lentiviral Vector

Brian C. Lewis,1,2,* Nachimuthu Chinnasamy,3 Richard A. Morgan,3 and Harold E. Varmus1,2

Cell Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10021,1 and Division of Basic Sciences, National Cancer Institute,2 and Clinical Gene Therapy Branch, National Human Genome Research Institute,3 Bethesda, Maryland 20892

Received 16 February 2001/Accepted 23 June 2001

We are using avian leukosis-sarcoma virus (ALSV) vectors to generate mouse tumor models in transgenic mice expressing TVA, the receptor for subgroup A ALSV. Like other classical retroviruses, ALSV requires cell division to establish a provirus after infection of host cells. In contrast, lentiviral vectors are capable of integrating their viral DNA into the genomes of nondividing cells. With the intention of initiating tumorigenesis in resting, TVA-positive cells, we have developed a system for the preparation of a human immunodeficiency virus type 1 (HIV-1)-based lentiviral vector, pseudotyped with the envelope protein of ALSV subgroup A (EnvA). The HIV(ALSV-A) vector retains the requirement for TVA on the surface of target cells and can be produced at titers of 5 × 103 infectious units (IU)/ml. By inserting the central polypurine tract (cPPT) from the HIV-1 pol gene and removing the cytoplasmic tail of EnvA, the pseudotype can be produced at titers approaching 105 IU/ml and can be concentrated by ultracentrifugation to titers of 107 IU/ml. HIV(ALSV-A) also infects embryonic fibroblasts derived from transgenic mice in which TVA expression is driven by the beta -actin promoter. In addition, this lentivirus pseudotype efficiently infects these fibroblasts after cell cycle arrest, when they are resistant to infection by ALSV vectors. This system may be useful for introducing genes into somatic cells in adult TVA transgenic animals and allows evaluation of the effects of altered gene expression in differentiated cell types in vivo.

* Corresponding author. Mailing address: Memorial Sloan-Kettering Cancer Center, 1275 York Ave., Box 62, New York, NY 10021. Phone: (212) 639-6362. Fax: (212) 717-3125. E-mail: lewisb{at}mskcc.org.

Journal of Virology, October 2001, p. 9339-9344, Vol. 75, No. 19
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.19.9339-9344.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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