Use of a herpes thymidine kinase/neomycin phosphotransferase chimeric gene for metabolic suicide gene transfer

Fabio Candotti, Riad Agbaria, Craig A. Mullen, Renaud Touraine, Jan Balzarini, David G. Johns, R. Michael Blaese

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Metabolic suicide gene transfer is widely applied for gene therapy of cancer, and retroviral vectors expressing the herpes simplex virus thymidine kinase (HSV-tk) gene are commonly used in clinical trials. Most of these vectors contain positive selectable markers that undoubtedly facilitate the determination of viral titer and the identification of high-titer producer clones. However, the presence of additional transcriptional units may result in reduced expression of the gene of interest. The use of fusion genes expressing bifunctional proteins may help to overcome this problem. We have constructed a retroviral vector carrying the TNFUS69 chimeric gene, which originates from the fusion of the HSV-tk and neomycin phosphotransferase II genes, and evaluated the functional expression of the encoded fusion protein. In vitro, expression of the fusion gene conferred to target cells both resistance to neomycin and selective sensitivity to the antiherpetic drugs ganciclovir and (E)-5-(2-bromovinyl)-2'-deoxyuridine. Cells transduced with the fusion gene, however, showed reduced ability to phosphorylate ganciclovir compared with cells expressing the native HSV-tk. Therefore, although the fusion gene may be used as a constituent of retroviral cassettes for positive and negative selection in vitro, its usefulness for suicide gene transfer applications in vivo may depend upon the possibility of using (E)-5-(2- bromovinyl)-2'-deoxyuridine in a clinical context.

Original languageEnglish
Pages (from-to)574-580
Number of pages7
JournalCancer Gene Therapy
Volume7
Issue number4
DOIs
StatePublished - 1 Jan 2000
Externally publishedYes

Keywords

  • (E)-5-(2- bromovinyl)-2'-deoxyuridine
  • Ganciclovir
  • Gene therapy
  • Retroviral vector
  • Suicide gene

Fingerprint

Dive into the research topics of 'Use of a herpes thymidine kinase/neomycin phosphotransferase chimeric gene for metabolic suicide gene transfer'. Together they form a unique fingerprint.

Cite this