Generation of helper-free amphotropic retroviruses that transduce a dominant-acting, methotrexate-resistant dihydrofolate reductase gene

We constructed several retroviruses which transduced a mutant dihydrofolate reductase gene that was resistant to methotrexate inhibition and functioned as a dominant selectable marker. The titer of dihydrofolate reductase-transducing virus produced by virus-producing cells could be increased to very high levels by selection of the cells in increasing concentrations of methotrexate. Helper virus-free dihydrofolate reductase-transducing virus was also generated by using a broad-host-range amphotropic retroviral packaging system. Cell lines producing helper-free dihydrofolate reductase-transducing virus with a titer of 4 X 10(6) per ml were generated. These retroviral vectors should have general utility for high-efficiency transduction of genes in cultured cells and in animals.

[1]  R. Evans,et al.  Infectious and selectable retrovirus containing an inducible rat growth hormone minigene. , 1984, Science.

[2]  I. Verma,et al.  Expression of a retrovirus encoding human HPRT in mice. , 1984, Science.

[3]  A. Efstratiadis,et al.  Cell-specified expression of a selectable hybrid gene. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[4]  E. Gilboa,et al.  Expression of genes introduced into cells by retroviral infection is more efficient than that of genes introduced into cells by DNA transfection , 1984, Journal of virology.

[5]  H. Temin,et al.  Expression of complete chicken thymidine kinase gene inserted in a retrovirus vector , 1984, Molecular and cellular biology.

[6]  I. Verma,et al.  Two base changes restore infectivity to a noninfectious molecular clone of Moloney murine leukemia virus (pMLV-1) , 1984, Journal of virology.

[7]  P. Jolicoeur,et al.  Physical mapping of the Fv-1 tropism host range determinant of BALB/c murine leukemia viruses , 1983, Journal of virology.

[8]  A. Joyner,et al.  Retrovirus transduction: generation of infectious retroviruses expressing dominant and selectable genes is associated with in vivo recombination and deletion events , 1983, Molecular and cellular biology.

[9]  H. Temin,et al.  Construction of a helper cell line for avian reticuloendotheliosis virus cloning vectors , 1983, Molecular and cellular biology.

[10]  I. Verma,et al.  A transmissible retrovirus expressing human hypoxanthine phosphoribosyltransferase (HPRT): gene transfer into cells obtained from humans deficient in HPRT. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[11]  J. Bertino,et al.  Genetic transformation of murine bone marrow cells to methotrexate resistance. , 1983, Blood.

[12]  I. Weinstein,et al.  Design of a Retrovirus-Derived Vector for Expression and Transduction of Exogenous Genes in Mammalian Cells , 1983, Molecular and cellular biology.

[13]  A. Levinson,et al.  Isolation and expression of an altered mouse dihydrofolate reductase cDNA. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[14]  D. Baltimore,et al.  Construction of a retrovirus packaging mutant and its use to produce helper-free defective retrovirus , 1983, Cell.

[15]  R. Weinberg,et al.  Adaptation of a retrovirus as a eucaryotic vector transmitting the herpes simplex virus thymidine kinase gene , 1982, Molecular and cellular biology.

[16]  I. Verma,et al.  Nucleotide sequence of the genome of a murine sarcoma virus , 1981, Cell.

[17]  R. Lerner,et al.  Nucleotide sequence of Moloney murine leukaemia virus , 1981, Nature.

[18]  H. Temin,et al.  Formation of infectious progeny virus after insertion of herpes simplex thymidine kinase gene into DNA of an avian retrovirus , 1981, Cell.

[19]  D. Haber,et al.  Properties of an altered dihydrofolate reductase encoded by amplified genes in cultured mouse fibroblasts. , 1981, The Journal of biological chemistry.

[20]  E. Scolnick,et al.  Construction and isolation of a transmissible retrovirus containing the src gene of Harvey murine sarcoma virus and the thymidine kinase gene of herpes simplex virus type 1 , 1981, Journal of virology.

[21]  M. Pearson,et al.  Enhancing the efficiency of DNA-mediated gene transfer in mammalian cells , 1981, Somatic cell genetics.

[22]  D. Lowy,et al.  Genomes of murine leukemia viruses isolated from wild mice , 1981, Journal of virology.

[23]  R. Schilsky,et al.  Clinical pharmacology of methotrexate. , 1981, Cancer treatment reports.

[24]  W. Salser,et al.  Gene transfer in intact animals , 1980, Nature.

[25]  E. Canaani,et al.  The transforming gene of Moloney murine sarcoma virus , 1979, Nature.

[26]  Tom Maniatis,et al.  Transformation of mammalian cells with genes from procaryotes and eucaryotes , 1979, Cell.

[27]  R T Schimke,et al.  Gene amplification and drug resistance in cultured murine cells. , 1978, Science.

[28]  W. S. Hayward,et al.  An avian oncovirus mutant (SE 21Q1b) deficient in genomic RNA: Biological and biochemical characterization , 1978, Cell.

[29]  J. Bertino,et al.  Selective multiplication of dihydrofolate reductase genes in methotrexate-resistant variants of cultured murine cells. , 1978, The Journal of biological chemistry.

[30]  R. Eckner,et al.  Defective Friend Spleen Focus-Forming Virus: Interfering Properties and Isolation Free from Standard Leukemia-Inducing Helper Virus , 1977, Journal of virology.

[31]  D. Bolognesi,et al.  A novel murine oncornavirus with dual eco- and xenotropic properties. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[32]  W. Nyhan,et al.  Reversion in expression of hypoxanthine-guanine phosphoribosyl transferase following cell hybridization. , 1975, Journal of cell science.

[33]  A. van der Eb,et al.  A new technique for the assay of infectivity of human adenovirus 5 DNA. , 1973, Virology.

[34]  J. Hartley,et al.  Plaque assay techniques for murine leukemia viruses. , 1970, Virology.