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. 1997 Nov 1;25(21):4416–4418. doi: 10.1093/nar/25.21.4416

Transient gene expression from yeast artificial chromosome DNA in mammalian cells is enhanced by adenovirus.

M Chen 1, S T Compton 1, V F Coviello 1, E D Green 1, M A Ashlock 1
PMCID: PMC147053  PMID: 9336477

Abstract

The introduction of high molecular weight DNA into mammalian cells is useful for gene expression studies. However, current transfection strategies are inefficient, necessitating propagation of stable DNA transformants prior to analysis of gene expression. Here we demonstrate that transient lipid-mediated DNA transfection can be used to assess gene expression from yeast artificial chromosomes (YACs) containing the 230 kb cystic fibrosis transmembrane conductance regulator gene ( CFTR ) and Escherichia coli lacZ . We also show that psoralen-UV inactivated adenovirus significantly enhances transfection efficiency. The ability to deliver high molecular weight DNA using lipid-mediated transfection should expedite the analysis of large human genes contained within artificial chromosome vectors.

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Selected References

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  1. Anand R., Ogilvie D. J., Butler R., Riley J. H., Finniear R. S., Powell S. J., Smith J. C., Markham A. F. A yeast artificial chromosome contig encompassing the cystic fibrosis locus. Genomics. 1991 Jan;9(1):124–130. doi: 10.1016/0888-7543(91)90229-8. [DOI] [PubMed] [Google Scholar]
  2. Baker A., Cotten M. Delivery of bacterial artificial chromosomes into mammalian cells with psoralen-inactivated adenovirus carrier. Nucleic Acids Res. 1997 May 15;25(10):1950–1956. doi: 10.1093/nar/25.10.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bellett A. J., Li P., David E. T., Mackey E. J., Braithwaite A. W., Cutt J. R. Control functions of adenovirus transformation region E1A gene products in rat and human cells. Mol Cell Biol. 1985 Aug;5(8):1933–1939. doi: 10.1128/mcb.5.8.1933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bergelson J. M., Cunningham J. A., Droguett G., Kurt-Jones E. A., Krithivas A., Hong J. S., Horwitz M. S., Crowell R. L., Finberg R. W. Isolation of a common receptor for Coxsackie B viruses and adenoviruses 2 and 5. Science. 1997 Feb 28;275(5304):1320–1323. doi: 10.1126/science.275.5304.1320. [DOI] [PubMed] [Google Scholar]
  5. Cotten M., Wagner E., Zatloukal K., Phillips S., Curiel D. T., Birnstiel M. L. High-efficiency receptor-mediated delivery of small and large (48 kilobase gene constructs using the endosome-disruption activity of defective or chemically inactivated adenovirus particles. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6094–6098. doi: 10.1073/pnas.89.13.6094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fernandez-Luna J. L., Matthews R. J., Brownstein B. H., Schreiber R. D., Thomas M. L. Characterization and expression of the human leukocyte-common antigen (CD45) gene contained in yeast artificial chromosomes. Genomics. 1991 Jul;10(3):756–764. doi: 10.1016/0888-7543(91)90460-v. [DOI] [PubMed] [Google Scholar]
  7. Gnirke A., Huxley C., Peterson K., Olson M. V. Microinjection of intact 200- to 500-kb fragments of YAC DNA into mammalian cells. Genomics. 1993 Mar;15(3):659–667. doi: 10.1006/geno.1993.1121. [DOI] [PubMed] [Google Scholar]
  8. Harrington J. J., Van Bokkelen G., Mays R. W., Gustashaw K., Willard H. F. Formation of de novo centromeres and construction of first-generation human artificial microchromosomes. Nat Genet. 1997 Apr;15(4):345–355. doi: 10.1038/ng0497-345. [DOI] [PubMed] [Google Scholar]
  9. Heard E., Kress C., Mongelard F., Courtier B., Rougeulle C., Ashworth A., Vourc'h C., Babinet C., Avner P. Transgenic mice carrying an Xist-containing YAC. Hum Mol Genet. 1996 Apr;5(4):441–450. doi: 10.1093/hmg/5.4.441. [DOI] [PubMed] [Google Scholar]
  10. Huxley C. Transfer of YACs to mammalian cells and transgenic mice. Genet Eng (N Y) 1994;16:65–91. [PubMed] [Google Scholar]
  11. Mastrangeli A., Danel C., Rosenfeld M. A., Stratford-Perricaudet L., Perricaudet M., Pavirani A., Lecocq J. P., Crystal R. G. Diversity of airway epithelial cell targets for in vivo recombinant adenovirus-mediated gene transfer. J Clin Invest. 1993 Jan;91(1):225–234. doi: 10.1172/JCI116175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mogayzel P. J., Jr, Henning K. A., Bittner M. L., Novotny E. A., Schwiebert E. M., Guggino W. B., Jiang Y., Rosenfeld M. A. Functional human CFTR produced by stable Chinese hamster ovary cell lines derived using yeast artificial chromosomes. Hum Mol Genet. 1997 Jan;6(1):59–68. doi: 10.1093/hmg/6.1.59. [DOI] [PubMed] [Google Scholar]
  13. Rosenfeld M. A. Human artificial chromosomes get real. Nat Genet. 1997 Apr;15(4):333–335. doi: 10.1038/ng0497-333. [DOI] [PubMed] [Google Scholar]
  14. Rosenfeld M. A., Yoshimura K., Trapnell B. C., Yoneyama K., Rosenthal E. R., Dalemans W., Fukayama M., Bargon J., Stier L. E., Stratford-Perricaudet L. In vivo transfer of the human cystic fibrosis transmembrane conductance regulator gene to the airway epithelium. Cell. 1992 Jan 10;68(1):143–155. doi: 10.1016/0092-8674(92)90213-v. [DOI] [PubMed] [Google Scholar]
  15. Setoguchi Y., Danel C., Crystal R. G. Stimulation of erythropoiesis by in vivo gene therapy: physiologic consequences of transfer of the human erythropoietin gene to experimental animals using an adenovirus vector. Blood. 1994 Nov 1;84(9):2946–2953. [PubMed] [Google Scholar]
  16. Yoshimura K., Rosenfeld M. A., Seth P., Crystal R. G. Adenovirus-mediated augmentation of cell transfection with unmodified plasmid vectors. J Biol Chem. 1993 Feb 5;268(4):2300–2303. [PubMed] [Google Scholar]

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