High efficiency gene transfer into mammalian cells.

We have generalized the protocol of gene transfer, greatly increasing the variety of cells that can be used as recipients of foreign genes. Our approach has been to use a transient assay system that allows rapid screening of expression of foreign DNA. When the initial steps of gene transfer have been optimized with the transient system, these defined conditions are used to yield efficient stable transformation. We have seen that primate cells, including human cells, can be used in gene transfer experiments at levels sensitive enough to allow detection of single copy gene function. Recently we have also used this approach successfully with undifferentiated embryonic cells.

[1]  Hans R. Schöler,et al.  Specific interaction between enhancer-containing molecules and cellular components , 1984, Cell.

[2]  B. Howard,et al.  Expression of recombinant plasmids in mammalian cells is enhanced by sodium butyrate. , 1983, Nucleic acids research.

[3]  B. Howard,et al.  High efficiency DNA-mediated transformation of primate cells. , 1983, Science.

[4]  P. Gruss,et al.  Multiple point mutations affecting the simian virus 40 enhancer. , 1983, Science.

[5]  I. Pastan,et al.  The Rous sarcoma virus long terminal repeat is a strong promoter when introduced into a variety of eukaryotic cells by DNA-mediated transfection. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[6]  P. Gruss,et al.  Host-specific activation of transcription by tandem repeats from simian virus 40 and Moloney murine sarcoma virus. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[7]  B. Howard,et al.  Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells , 1982, Molecular and cellular biology.

[8]  E. Lane,et al.  A rapid antibody assay system for screening hybridoma cultures. , 1981, Journal of immunological methods.

[9]  J. Banerji,et al.  Expression of a β-globin gene is enhanced by remote SV40 DNA sequences , 1981, Cell.

[10]  D. Pim,et al.  Monoclonal antibodies specific for simian virus 40 tumor antigens , 1981, Journal of virology.

[11]  I. Pastan,et al.  Identification of a functional promoter in the long terminal repeat of Rous sarcoma virus , 1980, Cell.

[12]  R. Mulligan,et al.  Expression of a bacterial gene in mammalian cells. , 1980, Science.

[13]  M. Wigler,et al.  The transfer and stable integration of the HSV thymidine kinase gene into mouse cells , 1978, Cell.

[14]  Richard Axel,et al.  Transfer of purified herpes virus thymidine kinase gene to cultured mouse cells , 1977, Cell.

[15]  P. Gruss,et al.  A Novel System Using the Expression of Chloramphenicol Acetyltransferase in Eukaryotic Cells Allows the Quantitative Study of Promoter Elements , 1984 .

[16]  T. Shenk,et al.  Enhancers and eukaryotic gene expression , 1983 .

[17]  P Berg,et al.  Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. , 1982, Journal of molecular and applied genetics.

[18]  W. V. Shaw.  [57] Chloramphenicol acetyltransferase from chloramphenicol-resistant bacteria , 1975 .