Abstract
Embryonal carcinoma (EC) cells can be efficiently transfected with cloned DNAs but there is a strong tendency for expression from transfected genes to be lost from stably transformed cells. To investigate the mechanism responsible for this loss of expression, we transfected P19 EC cells with a gene encoding theE. coli β-galactosidase and examined expression of this gene in clonal populations of cells. Cells that carry and express the β-galactosidase gene give rise to cells that do not express at a rate of about 0.02 events per cell per cell division. These non-expressing cells were of two types, some had lost the transfected genes while others had inactivated them. In those cells that retained but inactivated the transfected genes, the inactive state was stable and suppression was at the level of transcription initiation but not associated with increased DNA methylation. Because transfected DNAs integrate into the genome as tandem arrays, the gene loss and inactivation seen in EC cells may be analogous to the repeat-induced gene inactivation seen in lower eukaryotes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Literature Cited
Bell, J.C., Jardine, K., and McBurney, M.W. 1986.Mol. Cell. Biol. 6 2:617–625.
Skerjanc, I.S., Slack, R.S., and McBurney, M.W. 1994Mol. Cell. Biol. 14:8451–8459.
Slack, R.S., Sherjanc, I.S., Lach, B., Cratg, J., Jardine, K., and McBurney, M.W. 1995.J. Cell Biol. 129:779–788.
McBurney, M.W., Fournier, S., Schmidt-Kastner, P.K., Jardine, K., and Craig, J. 1995Somatic Cell Mol. Genet. 20:529–540.
McBurney, M.W. and Rogers, B.J. 1982Dev. Biol. 89:503–508.
Rudnicki, M.A. and McBurney, M.W. 1987. Teratocarcinomas and embryonic stem cells, a practical approach. E.J. Robertson, editor. IRL Press, Oxford. 19–49.
Chen, C. and Okayama, H. 1987Mol. Cell. Biol. 7:2745–2752.
Dannenberg, A.M.J. and Suga, M. 1981. Methods for Studying Mononuclear Phagocytes. S.O. Adams, P.J. Edelson, and M.S. Koren, editors. Academic Press, New York, 375–384.
Norton, P.A. and Coffin, J.M. 1985.Mol. Cell. Biol. 5:281–290.
Laird, P.W., Zidjerveld, A., Linders, K., Rudnicki, M.A., Jaenisch, R., and Berns, A. 1991.Nucleic Acids Res. 19:4293.
Sambrook, J., Fritsch, E.F., and Maniatis, T. 1989 Molecular Cloning, A Laboratory Manual. Harbor Laboratory Press. Cold Spring Harbor, New York.
Schibler, U., Hagenbuchle, O., Wellauer, P.K., and Pittet, A.C. 1983Cell 33:501–508.
McBurney, M.W., Fournier, S., Jardine, K., and Sutherland, L. 1995Somatic Cell Mol Genet. 20:515–528.
Vara, J.A., Portela, A., Ortin, J., and Jimenez, A. 1986Nucleic Acids Res. 14:4617–4624.
Adra, C.N., Boer, P.H., and McBurney, M.W. 1987Gene 60:65–74.
Boer, P.H., Potten, H., Adra, C.N., Jardine, K., Mullhofer, G., and McBurney, M.W. 1990Biochem. Genet. 28:299–308.
McBurney, M.W., Sutherland, L.C., Adra, C.N., Leclair, B., Rudnicki, M.A., and Jardine, K. 1991Nucleic Acids Res. 19:5755–5761.
Bartlett, M.H., Adra, C.N., Park, J., Chapman, V.M., and McBurney, M.W. 1991Somatic Cell Mol. Genet. 17:35–47.
Sutherland, L.C., St-Arnaud, R., and McBurney, M.W. 1995.Gene Expr. 4:265–279.
Eckner, R., Ewen, M.E., Newsome, D., Gerdes, M., DeCaprio, J.A., Lawrence, J.B., and Livingston, D.M. 1994Genes Dev. 8:869–884.
Kwok, R.P.S., Lundblad, J.R., Chrivia, J.C., Richards, J.P., Bächinger, H.P., Brennan, R.G., Roberts, S.G.E., Green, M.R., and Goodman, R.H. 1994Nature 370:223–226.
Khavari, P.A., Peterson, C.L., Tamkun, J.W., Mendel, D.B., and Crabtree, G.R. 1993.Nature 366: 170–174.
Bernards, R., Schackleford, G.M., Gerber, M.R., Horowitz, J.M., Friend, S.H., Schartl, M., Bogenmann, E., Rapaport, J.M., McGee, T., Dryja, T.P., and Weinberg, R.A. 1989Proc. Natl. Acad. Sci. U.S.A. 86:6474–6478.
Szyf, M., Tanigawa, G., and McCarthy, P.L., Jr. 1990Mol. Cell. Biol. 10:4396–4400.
Wigler, M., Sweet, R., Sim, G.K., Wold, B., Pellicer, A., Lacy, E., Maniatis, T., Silverstein, S., and Axel, R. 1977Cell 16:777–785.
Judelson, H.S., and Whittaker, S.L. 1995Curr. Genet. 28:571–579.
Singer, M.J., and Selker, E.U. 1995Curr. Top. Microbiol. Immunol. 197:165–178.
Flavell, R.B. 1994Proc. Natl. Acad. Sci. U.S.A. 91:3490–6X.
Assaad, F.F., and Signer, E.R. 1992Genetics 132:553–66X.
Dorer, D.R., and Henikoff, S. 1994Cell 77:993–1002.
Pawlik, K.M., Sun, C.W., Higgins, N.P., and Townes, T.M. 1995Gene 165:173–181.
Woods, C., LeFeuvre, C., Stewart, N., and Bacchetti, S. 1994Oncogene 9:2943–2950.
Gargano, S., Wang, P., Rusanganwa, E., and Bacchetti, S. 1995Mol. Cell. Biol. 15:6256–6261.
Bailey, A.D., Li, Z.J., Pavelitz, T., and Weiner, A.M. 1995Mol. Cell. Biol. 15:6246–6255.
Bootsma, D., and Hoeijmakers, J.H.J., 1993Nature 363:114–115.
Kennedy, B.K., Austriaco, N.R., Jr., Zhang, J., and Guarente, L. 1995Cell 80:485–496.
Smeal, T., James, C., Kennedy, B., Cole, F., and Guarente, L. 1996Cell 84:633–642.
Dimri, G.P., Lee, X.H., Basile, G., Acosta, M., Scott, C., Roskelley, C., Medrano, E.E., Linskens, M., Rubelj, I., Pereira-Smith, O., Peacocke, M., and Campisi, J. 1995Proc. Natl. Acad. Sci. U.S.A. 92:9363–9367.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Schmidt-Kastner, P.K., Jardine, K., Cormier, M. et al. Genes transfected into embryonal carcinoma stem cells are both lost and inactivated at high frequency. Somat Cell Mol Genet 22, 383–392 (1996). https://doi.org/10.1007/BF02369894
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02369894