Inducible, High-Level Production of Infectious Murine Leukemia Retroviral Vector Particles Pseudotyped with Vesicular Stomatitis Virus G Envelope Protein

Yanping Yang, Elio F. Vanin, Michael Whitt, Maarten Fornerod, Ronald Zwart, Richard D. Schneiderman, Gerard Grosveld, Arthur W. Nienhuis

Research output: Contribution to journalArticle

107 Citations (Scopus)

Abstract

Murine leukemia viruses (MuLV) have been adapted for use as gene transfer vectors for experimental and human gene therapy applications. Their utility for these purposes has been circumscribed by the limited host range and relatively low titer of available producer clones. Pseudotyping of MuLV particles with the vesicular stomatitis virus envelope protein (VSV-G), expressed transiently in cells producing MuLV Gag and Pol proteins, has yielded vector preparations with a broader host range that can be concentrated by ultracentrifugation. We have explored the use of steroid-inducible and tetracycline-modulated promoter systems (necessary because the VSV-G protein is toxic to cells when constitutively expressed) to derive stable producer cell lines capable of substantial production of VSV-G pseudotyped MuLV particles. A packaging cell line and producer clones capable of expressing a chimeric transcription factor, composed of the tetracycline repressor (tetR) and the VP16 trans-activating sequences of herpes simplex virus VP16 gene and containing the VSV-G coding sequences linked to a minimal promoter having seven tandem copies of the tetracycline responsive operator (tet°), exhibited high levels of VSV-G protein expression when cultured in the absence of tetracycline. Vector particles, produced at titers of 105–106 infectious colony forming units per ml (cfu/ml), could be concentrated effectively by ultracentrifugation yielding vector preparations having a titer of 109 cfu/ml. These cell lines grew normally when VSV-G protein expression was repressed by tetracycline. Such producer clones hold promise for future human gene therapy applications.

Original languageEnglish (US)
Pages (from-to)1203-1213
Number of pages11
JournalHuman Gene Therapy
Volume6
Issue number9
DOIs
StatePublished - Sep 1 1995

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Viral Envelope Proteins
Tetracycline
Murine Leukemia Viruses
Leukemia
GTP-Binding Proteins
Clone Cells
Host Specificity
Ultracentrifugation
Cell Line
Genetic Therapy
Virion
gag-pol Fusion Proteins
Stem Cells
Vesicular Stomatitis
Poisons
Product Packaging
Simplexvirus
Genes
Transcription Factors
Steroids

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Inducible, High-Level Production of Infectious Murine Leukemia Retroviral Vector Particles Pseudotyped with Vesicular Stomatitis Virus G Envelope Protein. / Yang, Yanping; Vanin, Elio F.; Whitt, Michael; Fornerod, Maarten; Zwart, Ronald; Schneiderman, Richard D.; Grosveld, Gerard; Nienhuis, Arthur W.

In: Human Gene Therapy, Vol. 6, No. 9, 01.09.1995, p. 1203-1213.

Research output: Contribution to journalArticle

Yang, Yanping ; Vanin, Elio F. ; Whitt, Michael ; Fornerod, Maarten ; Zwart, Ronald ; Schneiderman, Richard D. ; Grosveld, Gerard ; Nienhuis, Arthur W. / Inducible, High-Level Production of Infectious Murine Leukemia Retroviral Vector Particles Pseudotyped with Vesicular Stomatitis Virus G Envelope Protein. In: Human Gene Therapy. 1995 ; Vol. 6, No. 9. pp. 1203-1213.
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