Amplified and tissue-directed expression of retroviral vectors using ping-pong techniques

Maureen Hoatlin, S. L. Kozak, C. Spiro, David Kabat

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Ping-pong amplification is an efficient process by which helper-free retrovirions replicate in cocultures of cell lines that package retroviruses into distinct hostrange envelopes [11]. Transfection of a retroviral vector DNA into these cocultures results in massive virus production, with potentially endless cross-infection between different types of packaging cells. Because the helperfree virus spreads efficiently throughout the coculture, it is unnecessary to use dominant selectable marker genes, and the retroviral vectors can be simplified and optimized for expressing a single gene of interest. The most efficient ping-pong vector, pSFF, derived from the Friend erythroleukemia virus, has been used for high-level expression of several genes that could not be expressed with commonly employed two-gene retroviral vectors. Contrary to previous claims, problems of vector recombination are not inherent to ping-pong methods. Indeed, the pSFF vector has not formed replication-competent recombinants as shown by stringent assays. Here we review these methods, characterize the ping-pong process using the human erythropoietin gene as a model, and describe a new vector (pSFY) designed for enhanced expression in T lymphocytes. Factors that limit tissue-specific expression are reviewed.

Original languageEnglish (US)
Pages (from-to)113-120
Number of pages8
JournalJournal of Molecular Medicine
Volume73
Issue number3
DOIs
StatePublished - Mar 1995

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Coculture Techniques
Genes
Friend murine leukemia virus
Viruses
Leukemia, Erythroblastic, Acute
Thromboplastin
Product Packaging
Retroviridae
Cross Infection
Erythropoietin
Genetic Recombination
Transfection
T-Lymphocytes
Gene Expression
Cell Line
DNA

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Amplified and tissue-directed expression of retroviral vectors using ping-pong techniques. / Hoatlin, Maureen; Kozak, S. L.; Spiro, C.; Kabat, David.

In: Journal of Molecular Medicine, Vol. 73, No. 3, 03.1995, p. 113-120.

Research output: Contribution to journalArticle

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