title: In vivo drug-selectable markers in gene therapy.
abstract: Two-gene vectors with positive or positive-negative drug-selectable markers enable the expansion or elimination of gentetically modified cells in vivo. We have established a bicistronic retroviral vector system which utilizes an internal ribosome entry site (IRES) to co-express two independent genes with high efficiency. As a positive-negative (suicide) marker, Herpes simplex virus thymidine kinase was co-expressed with the human multidrug resistance gene, MDR1. Using this vector, almost all the MDR1-transduced cells showed hypersensitivity to a nucleoside analog, ganciclovir. As a dominant selectable marker, the MDR1 gene was co-expressed with alpha-galactosidase A for the model of gene therapy of Fabry disease. Vincristine selection efficiently enhanced the population of transduced cells expressing the second non-selectable genes. These drug-selectable retroviral vectors could be applicable to the therapy of many diseases.
authors.affiliation: Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, Tokyo, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Sugimoto
authors.forename: T
authors.initials: T
authors.lastname: Tsuruo
mesh_terms: D016475:3T3 Cells; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000096:biosynthesis; D000818:Animals; D018432:Drug Resistance, Multiple / Q000235:genetics*; D030342:Genetic Diseases, Inborn / Q000628:therapy; D005819:Genetic Markers; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006251:Harvey murine sarcoma virus; D006801:Humans; D051379:Mice; D012091:Repetitive Sequences, Nucleic Acid; D012190:Retroviridae; D012270:Ribosomes / Q000378:metabolism; D018139:Simplexvirus / Q000201:enzymology / Q000235:genetics; D013937:Thymidine Kinase / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection / Q000379:methods; D000519:alpha-Galactosidase / Q000096:biosynthesis / Q000235:genetics
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D005819:Genetic Markers; D013937:Thymidine Kinase; D000519:alpha-Galactosidase