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Efficient Expression of Drug-selectable Genes in Retroviral Vectors Under Control of an Internal Ribosome Entry Site

Bio/Technology volume 12, pages 694–698 (1994)Cite this article

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 hyper-sensitivity 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.

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Author notes

  1. Yoshikazu Sugimoto

    Present address: Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, 1-37-1 Kami-Ikebukuro, Toshima-ku, Tokyo, 170, Japan

Authors and Affiliations

  1. National Cancer Institute, Laboratory of Cell Biology, National Institute of Health, Bethesda, MD, 20892

    Yoshikazu Sugimoto, Ivan Aksentijevich & Michael M. Gottesman

  2. National Cancer Institute, Laboratory of Molecular Biology, National Institute of Health, Bethesda, MD, 20892

    Ira Pastan

Authors
  1. Yoshikazu Sugimoto
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  2. Ivan Aksentijevich
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  3. Michael M. Gottesman
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  4. Ira Pastan
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Correspondence to Ira Pastan.

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Sugimoto, Y., Aksentijevich, I., Gottesman, M. et al. Efficient Expression of Drug-selectable Genes in Retroviral Vectors Under Control of an Internal Ribosome Entry Site. Nat Biotechnol 12, 694–698 (1994). https://doi.org/10.1038/nbt0794-694

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