Efficient expression of drug-selectable genes in retroviral vectors under control of an internal ribosome entry site

Yoshikazu Sugimoto, Ivan Aksentijevich, Michael M. Gottesman, Ira Pastan

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

We describe a new retroviral vector system pSXLC/pHa that utilizes a putative internal ribosome entry site (IRES) from encephalomyocarditis virus downstream from a multicloning site to co-express drug-selectable markers with a second non-selectable cDNA in a eukaryotic expression vector. The positive drug-selectable marker, MDR1, and the positive-negative marker, herpes simplex virus thymidine kinase (HSV-TK), were successfully introduced and expressed in the pSXLC/pHa system. The pSXLC-MDR and pSXLC-TK vectors contain the drug-selectable genes under translational control of the IRES and multiple cloning sites upstream for insertion of second cDNAs which can be co-expressed in this system. The inserts of these pSXLC plasmids were designed for easy transfer to the pHa retrovirus vector which has a strong promoter from Harvey murine sarcoma virus. The IRES-MDR-carrying retroviral vector, pHa-MCS-IRES-MDR, conferred resistance to vincristine and adriamycin. The IRES-TK-containing vector, pHa-MCS-IRES-TK conferred HAT-resistance in TK-deficient cells and the transfectants showed hypersensitivity to ganciclovir. These "flexible" vectors should be useful for co-expression of genes for selectable gene transfer and for positive-negative (suicide) selections in vitro and in vivo.

Original languageEnglish
Pages (from-to)694-698
Number of pages5
JournalNature Biotechnology
Volume12
Issue number7
Publication statusPublished - 1994
Externally publishedYes

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Genes
Pharmaceutical Preparations
Viruses
Harvey murine sarcoma virus
Complementary DNA
Encephalomyocarditis virus
Ganciclovir
Thymidine Kinase
Gene transfer
Vincristine
Simplexvirus
Retroviridae
Cloning
Doxorubicin
Suicide
Internal Ribosome Entry Sites
Organism Cloning
Hypersensitivity
Plasmids
Gene Expression

ASJC Scopus subject areas

  • Microbiology
  • Biotechnology

Cite this

Efficient expression of drug-selectable genes in retroviral vectors under control of an internal ribosome entry site. / Sugimoto, Yoshikazu; Aksentijevich, Ivan; Gottesman, Michael M.; Pastan, Ira.

In: Nature Biotechnology, Vol. 12, No. 7, 1994, p. 694-698.

Research output: Contribution to journalArticle

Sugimoto, Yoshikazu ; Aksentijevich, Ivan ; Gottesman, Michael M. ; Pastan, Ira. / Efficient expression of drug-selectable genes in retroviral vectors under control of an internal ribosome entry site. In: Nature Biotechnology. 1994 ; Vol. 12, No. 7. pp. 694-698.
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