Abstract
The introduction of exogenous DNA into mammalian cells can ultimately result in the integration of the transfected DNA into the host genome (1,2). Any genes containing appropriate expression signals will, in most cases, be expressed both prior to and after integration. However, stable expression of the transfected genes is not always observed (3,4). One reason this is believed to occur is because the site of integration of the foreign DNA is virtually random with respect to both the exogenous DNA and the particular human chromosome (5). Therefore, it can be expected that not only the expression, but also the stability and copy number of the transfected gene will depend on the integration sites and chromosomal flanking sequences.
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References
Heartlein, M. W., Knoll, J. H. M., and Latt, S. A. (1988) Chromosome instability associated with human alphoid DNA transfected into the Chinese hamster genome. Mol. Cell. Biol. 8, 3611–3618.
Talarico, D., Peverali, A. F., Ginelli, E., Meneveri, R., Mondello, C., and Della Valle, G. (1988) Satellite DNA induces unstable expression of the adjacent herpes simplex virus tk gene cotransfected in mouse cells. Mol. Cell Biol. 8, 1336–1344.
Butner, K., and Lo, C. W. (1986) Modulation of tk expression in mouse pericentromeric heterochromatin. Mol. Cell. Biol. 6, 4440–4449.
Butner, K. A., and Lo, C. W. (1986) High frequency DNA rearrangements associated with mouse centromeric satellite DNA. J. Mol. Biol. 187, 547–556.
Murnane, J. P., Yezzi, M. J., and Young, B. R. (1990) Recombination events during integration of transfected DNA into normal human cells. Nucleic Acids Res. 18, 2733–2738.
Grigliatti, T. (1991) Position-effect variegation—an assay for nonhistone chromosomal proteins and chromatin assembly and modifying factors. Methods in Cell Biol. 35, 587–627.
Beridze, T. (1986) Satellite DNA. Springer-Verlag, Berlin, Germany.
Wallenburg, J. C., Nepveu, A., and Chartrand, P. (1987) Integration of a vector containing rodent repetitive elements in the rat genome. Nucleic Acids Res. 15, 7849–7863.
Fouquet, C. and DuBow, M. S. (1992) Effect of cis-located human satellite DNA on the electroporation efficiency of neo and HSV-1 tk containing plasmids. Mut. Res. 284, 321–328.
Mulligan, R. C., and Berg, P. (1980) Expression of a bacterial gene in mammalian cells. Science 209, 1422–1427.
Potter, H., Selden, R. F., and Kingston, R. E. (1987) Transfection by electroporation, in Current Protocols in Molecular Biology (Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A., and Struhl, K., eds.), Greene Publishing Associates and Wiley-Interscience, Brooklyn, NY, pp. 9.3.1–9.5.6.
Goring, D. R., and DuBow, M. S. (1985) A cytotoxic effect associated with 9-(1,3-dihydroxy-2-propoxymethyl) guanine is observed during the selection for drug resistant human cells containing a single Herpesvirus thymidine kinase gene. Biochem. Biophys. Res. Commun. 133, 195–201.
Sol, K., Lapointe, M., Macleod, M., Nadeau, C., and DuBow, M. S. (1986) A cloned fragment of Hela DNA containing consensus sequences of satellite II and III DNA hybridizes with the Drosophila P-element and with the 1.8 kb family of human KpnI fragments. Biochim. Biophys. Acta. 868, 128–135.
Rhim, J. S., Cho, H. Y., and Huebner, R. J. (1975) Non-producer human cells induced by murine sarcoma virus. Int. J. Cancer 15, 23–29.
Horita, A., and Weber, L. J. (1964) Skin penetrating property of drugs dissolved in dimethylsulfoxide (DMSO) and other vehicles. Life Sci. 3, 1389–1395.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Plasmid vectors, in Molecular Cloning, a Laboratory Manual, vol. 1, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp. 1.42–1.46.
Tolias, P. P., and DuBow, M. S. (1987) The amino terminus of the bacteriophage D108 transposase protein contains a two-component, sequence-specific, DNA-binding domain. Virology 157, 117–126.
Stubblefield, E. (1968) Synchronization methods for mammalian cell cultures, in Methods in Cell Physiology (Prescott, D. M., ed.), Academic, New York, pp. 25–43.
Chu, G., Hayakawa, H., and Berg P. (1987) Electroporation for the efficient transfection of mammalian cells with DNA. Nucleic Acids Res. 15, 1311–1326.
Potter, H., Weir, L., and Leder, P. (1984) Enhancer-dependent expression of human κ immunoglobulin genes introduced into mouse pre-B lymphocytes by electroporation. Proc. Natl. Acad. Sci. USA 81, 7161–7165.
Hartman, S. C. and Mulligan, R. C. (1988) Two dominant-acting selectable markers for gene transfer studies in mammalian cells. Proc. Natl. Acad. Sci. USA 85, 8047–8051.
Artlet, P., Grannemann, R., Friel, J., Bartsch, J., and Hauser, H. (1991) The prokaryotic neomycin-resistance-encoding gene acts as a transcriptional silencer in eukaryotic cells. Gene 99, 249–254.
Brisebois, J. J. and DuBow, M. S. (1993) Selection for spontaneous null mutations in a chromosomally integrated HSV-1 thymidine kinase gene yields deletions and a mutation caused by intragenic illegitimate recombination. Mut. Res. 287, 191–205.
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Saint-Dic, D., DuBow, M.S. (1995). Effect of Cis-Located Human Satellite DNA on Electroporation Efficiency. In: Nickoloff, J.A. (eds) Animal Cell Electroporation and Electrofusion Protocols. Methods in Molecular Biology, vol 48. Humana Press. https://doi.org/10.1385/0-89603-304-X:199
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DOI: https://doi.org/10.1385/0-89603-304-X:199
Publisher Name: Humana Press
Print ISBN: 978-0-89603-304-7
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