Abstract
In order to identify regulatory elements that direct widespread in vivo expression of a linked gene, we have examined one of the human aldolase A alternative promoters, the ubiquitous pH promoter, which is active in most foetal and adult tissues. We have used the pH promoter region to drive expression of an heterologous CAT reporter gene in transgenic mice. We show that a short 820 bp pH promoter fragment is able to confer a ubiquitous and reproducible activity pattern on the CAT reporter gene in most of the transgenic lines analysed, with a particularly high level of expression in adult skeletal muscle. Activity of this transgene was detected from early embryonic stages. Therefore, this pH promoter region appears to be a powerful tool to direct ubiquitous and early expression of a transgene in vivo. Deletion analysis revealed that: (i) the region between −651 and −369 bp relative to the pH promoter transcription start site includes DNA elements capable of overriding effects of the surrounding chromatin at the integration site, (ii) the region between −285 and −211 bp is involved in pH promoter tissue-specific expression pattern in skeletal muscle and/or nervous tissues, (iii) the region located between −211 and −108 bp is necessary for its ubiquitous and muscle-pre dominant activity and (iv) the most proximal region downstream from −108 bp is still sufficient to confer an activity in brain and lung
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Al-Shawi, R., Kinnaird, J., Burke, J. and Bishop, J.O. (1990) Expression of a foreign gene in a line of transgenic mice is modulated by a chromosomal position effect. Mol. Cell. Biol. 10, 1192-8.
Bassel-Duby, R., Hernandez, M.D., Gonzalez, M.A., Krueger, J.K. and Williams, R.S. (1992) A 40-kilodalton protein binds specifically to an upstream sequence element essential for muscle-specific transcription of the human myoglobin promoter. Mol. Cell. Biol. 12, 5024-32.
Bassel-Duby, R., Hernandez, M.D., Yang, Q., Rochelle, J.M., Seldin, M.F. and Williams, R.S. (1994) Myocyte nuclear factor, a novel winged-helix transcription factor under both developmental and neural regulation in striated myocytes. Mol. Cell. Biol. 14, 4596-605.
Behringer, R.R., Cate, R.L., Froelick, G.J., Palmiter, R.D. and Brinster, R.L. (1990) Abnormal sexual development in transgenic mice chronically expressing Müllerian inhibiting substance. Nature 345, 167-70.
Betz, U.A.K., Voßhenrich, C.A.J., Rajewsky, K. and Müller, W. (1996) Bypass of lethality with mosaic mice generated by Cre-loxP-mediated recombination. Curr. Biol. 6, 1307-16.
Buckingham, M.E. (1994) Muscle: the regulation of myogenesis. Curr. Opin. Genet. Dev. 4, 745-51.
Cavard, C., Grimber, G., Dubois, N., Chasse, J.F., Bennoun, M., Minet-Thuriaux, M., Kamoun, P. and Briand, P. (1988) Correction of mouse ornithine transcarbamylase deficiency by gene transfer into the germ line. Nucl. Acids Res. 16, 2099-110.
Choi, T., Huang, M., Gorman, C. and Jaenisch, R. (1991) A generic intron increases gene expression in transgenic mice. Mol. Cell. Biol. 11, 3070-4.
Concordet, J.P., Maire, P., Kahn, A. and Daegelen, D. (1991) A ubiquitous enhancer shared by two promoters in the human aldolase A gene. Nucl. Acids Res. 19, 4173-80.
Concordet, J.P., Salminen, M., Demignon, J., Moch, C., Maire, P., Kahn, A. and Daegelen, D. (1993) An opportunistic promoter sharing regulatory sequences with either a muscle-specific or a ubiquitous promoter in the human aldolase A gene. Mol. Cell. Biol. 13, 9-17.
Epner, E., Kim, C.G. and Groudine, M. (1992) What does the locus control region control? Curr. Biol. 2, 262-4.
Furth, P.A., Hennighausen, L., Baker, C., Beatty, B. and Woychick, R. (1991) The variability in activity of the universally expressed human cytomegalovirus immediate early gene 1 enhancer/promoter in transgenic mice. Nucl. Acids Res. 19, 6205-8.
Gautron, S., Maire, P., Hakim, V. and Kahn, A. (1991) Regulation of the multiple promoters of the human aldolase A gene: response of its two ubiquitous promoters to agents promoting cell proliferation. Nucl. Acids Res. 19, 767-74.
Gu, H., Marth, J.D., Orban, P.C., Mossmann, H. and Rajewsky, K. (1994) Deletion of a DNA polymerase β gene segment in T cells using cell type-specific gene targeting. Science 265, 103-6.
Hogan, B., Costantini, F. and Lacy, E. (1986) Manipulating the Mouse Embryo: a Laboratory Manual. Cold Spring Harbor, NY; USA: Cold Spring Harbor Laboratory Press.
Izzo, P., Costanzo, P., Lupo, A., Rippa, E., Paolella, G. and Salvatore, F. (1988) Human aldolase A gene: structural organization and tissue-specific expression by multiple promoters and alternate mRNA processing. Eur. J. Biochem. 174, 569-78.
Kistner, A., Gossen, M., Zimmermann, F., Jerecic, J., Ullmer, C., Lübbert, H. and Bujard, H. (1996) Doxycycline-mediated quantitative and tissue-specific control of gene expression in transgenic mice. Proc. Natl Acad. Sci. USA 93, 10933-8.
Kitsukawa, T., Shimono, A., Kawakami, A., Kondoh, H. and Fujisawa, H. (1995) Overexpression of a membrane protein, neuropilin, in chimeric mice causes anomalies in the cardiovascular system, nervous system and limbs. Development 121, 4309-18.
Klebig, M.L., Wilkinson, J.E., Geisler, J.G. and Woychik, R.P. (1995) Ectopic expression of the agouti gene in transgenic mice causes obesity, features of type II diabetes, and yellow fur. Proc. Natl Acad. Sci. USA 92, 4728-32.
Leturque, A., Loizeau, M., Vaulont S., Salminen, M. and Girard, J. (1996) Improvement of insulin action in diabetic transgenic mice selectively overexpressing GLUT4 in skeletal muscle. Diabetes 45, 23-7.
Levy, M.Y., Barron, L.G., Meyer, K.B. and Szoka Jr, F.C. (1996) Characterization of plasmid DNA transfer into mouse skeletal muscle: evaluation of uptake mechanism, expression and secretion of gene products into blood. Gene Therapy 3, 201-11.
Luckow, B. and Schütz, G. (1987) CAT constructions with multiple unique restriction sites for the functional analysis of eukaryotic promoters and regulatory elements. Nucl. Acids Res. 15, 5490.
Maire, P., Gautron, S., Hakim, V., Gregori, C., Mennecier, F. and Kahn, A. (1987) Characterization of three optional promoters in the 5′ region of the human aldolase A gene. J. Mol. Biol. 197, 425-38.
McBurney, M.W., Staines, W.A., Boekelheide, K., Parry, D., Jardine, K. and Pickavance, L. (1994) Murine Pgk-1 promoter drives widespread but not uniform expression in transgenic mice. Dev. Dynamics 200, 278-93.
Mehtali, M., LeMeur, M. and Lathe, R. (1990) The methylation-free status of a housekeeping transgene is lost at high copy number. Gene 91, 179-84.
Mikkelsen, T.R., Chapman, B., Din, N., Ingerslev, J., Kristensen, P., Poulsen, K. and Hjorth, J.P. (1992) Expression of a cytomegalovirus IE-1-factor VIII cDNA hybrid gene in transgenic mice. Transgenic Res. 1, 164-9.
Moch, C., Kahn, A. and Daegelen, D. (1996) Independence and interdependence of the three human aldolase A promoters in transgenic mice. Gene Expr. 6, 1-14.
Nakayama, M., Stauffer, J., Cheng, J., Banerjee-Basu, S., Wawrousek, E. and Buonanno, A. (1996) Common core sequences are found in skeletal muscle slow-and fast-fiber-type-specific regulatory elements. Mol. Cell. Biol. 16, 2408-17.
Overbeek, P.A., Lai, S.P., Van Quill, K.R. and Westphal, H. (1986) Tissue-specific expression in transgenic mice of a fused gene containing RSV terminal sequences. Science 231, 1574-7.
Paldi, A., Deltour, L. and Jami, J. (1993) Cis effect of lacZ sequences in transgenic mice. Transgenic Res. 2, 325-9.
Palmiter, R.D., Norstedt, G., Gelinas, R.E., Hammer, R.E. and Brinster, R.L. (1983) Metallothionein-human GH fusion genes stimulate growth of mice. Science 222, 809-14.
Palmiter, R.D., Sandgren, E.P., Avarbock, M.R., Allen, D.D. and Brinster, R.L. (1991) Heterologous introns can enhance expression of transgenes in mice. Proc. Natl Acad. Sci. USA 88, 478-82.
Parmacek, M.S., Ip, H.S., Jung, F., Shen, T., Martin, J.F., Vora, A.J., Olson, E.N. and Leiden, J.M. (1994) A novel myogenic regulatory circuit controls slow/cardiac troponin C gene transcription in skeletal muscle. Mol. Cell. Biol. 14, 1870-85.
Rüther, U., Müller, W., Sumida, T., Tokuhisa, T., Rajewsky, K. and Wagner, E.F. (1988) c-fos expression interferes with thymus development in transgenic mice. Cell 53, 847-56.
Salminen, M., Maire, P., Concordet, J.P., Moch, C., Porteu, A., Kahn, A. and Daegelen, D. (1994) Fast-muscle-specific expression of human aldolase A transgenes. Mol. Cell. Biol. 14, 6797-808.
Sassi, H., Fromont-Racine, M., Grange, T. and Pictet, R. (1995) Tissue specificity of a glucocorticoid-dependent enhancer in transgenic mice. Proc. Natl Acad. Sci. USA 92, 7197-201.
Schmidt, E.V., Christoph, G., Zeller, R. and Leder, P. (1990) The cytomegalovirus enhancer: a pan-active control element in transgenic mice. Mol. Cell. Biol. 10, 4406-11.
Schorpp, M., Jäger, R., Schellander, K., Schenkel, J., Wagner, E.F., Weiher, H. and Angel, P. (1996) The human ubiquitin C promoter directs high ubiquitous expression of transgenes in mice. Nucl. Acids Res. 24, 1787-8.
Sternberg, E.A., Spizz, G., Perry, W.M., Vizard, D., Weil, T. and Olson, E.N. (1988) Identification of upstream and intragenic regulatory elements that confer cell-type-restricted and differentiation-specific expression on the muscle creatine kinase gene. Mol. Cell. Biol. 8, 2896-909.
Thierry, A.R., Lunardi-Iskandar, Y., Bryant, J.L., Rabinovich, P., Gallo, R.C. and Mahan, L.C. (1995) Systemic gene therapy: biodistribution and long-term expression of a transgene in mice. Proc. Natl Acad. Sci. USA 92, 9742-6.
Tuil, D., Soulez, M., Montarras, D., Pinset, C., Kahn, A. and Phan-Dinh-Tuy, F. (1993) Activation of gene expression via CArG boxes during myogenic differentiation. Exp. Cell Res. 205, 32-8.
Winston, J.H., Hong, L., Datta, S.K. and Kellems, R.E. (1996) An intron 1 regulatory region from the murine adenosine deaminase gene can activate heterologous promoters for ubiquitous expression in transgenic mice. Somat. Cell Mol. Genet. 22, 261-78.
Yutzey, K.E., Kline, R.L. and Konieczny, S.F. (1989) An internal regulatory element controls troponin I gene expression. Mol. Cell. Biol. 9, 1397-405.
Zhang, L., Eddy, A., Teng, Y.T., Fritzler, M., Kluppel, M., Melet, F. and Bernstein, A. (1995) An immunological renal disease in transgenic mice that overexpress Fli-1, a member of the ets family of transcription factor genes. Mol. Cell. Biol. 15, 6961-70.
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Moch, C., Spitz, F., Porteu, A. et al. The human pH aldolase A promoter directs widespread but muscle-predominant expression in transgenic mice. Transgenic Res 7, 113–121 (1998). https://doi.org/10.1023/A:1008820409079
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DOI: https://doi.org/10.1023/A:1008820409079