Abstract
The development of transgenic technology during recent years has allowed researchers to probe much more deeply than was previously possible into the molecular mechanisms influencing embryonic development. Transgenic procedures allow the transfer of a cloned gene into a host genome or the mutation of specific genomic sequences via targeting in embryonic stem (ES) cells (see Chapter 7). Most commonly, transgenic gene transfer experiments have been used to study the phenotypic effects caused by the misexpression or overexpression of a transgene (1–3), or to investigate the transcriptional mechanisms underlying developmental and tissue-specific gene regulation (4–6). The use of reporter genes in this latter application will be dealt with in this chapter.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Balling, R., Mutter, G., Gruss, P., and Kessel, M. (1989) Craniofacial abnormalities induced by ectopic expression of the homeobox gene Hox-1.1 in transgenic mice. Cell 58, 337–347.
Wolgemuth, D. J., Behringer, R. R., Mostoller, M. P., Brinster, R. L., and Palmiter, R. D. (1989) Transgenic mice overexpressing the mouse homeobox-containing gene Hox-1.4 exhibit abnormal gut development. Nature 337, 464–467.
Lufkin, T., Mark, M., Hart, C. P., Dollé, P., LeMeur, M., and Chambon, P. (1992) Homeotic transformation of the occipital bones of the skull by ectopic expression of a homeobox gene. Nature 359, 835–841.
Goring, D. R., Rossant, J., Clapoff, S., Breitman, M. L., and Tsui, L.-C. (1987) In situ detection of β-galactosidase in lenses of transgenic mice with a γ-Crystallin/lacZ gene. Science 235, 456–458.
Whiting, J., Marshall, H., Cook, M., Krumlauf, R., Rigby, P. W. J., Stott, D., and Allemann, R. K. (1991) Multiple spatially specific enhancers are required to reconstruct the pattern of Hox-2.6 gene expression. Genes Dev. 5, 2048–2059.
Yee, S.-P. and Rigby, P. W. J. (1993) The regulation of myogenin gene expression during the embryonic development of the mouse. Genes Dev. 7, 1277–1289.
Hogan, B., Beddington, R., Costantini, F., and Lacy, L. (eds.) (1995) Manipulating the Mouse Embryo: A Laboratory Manual 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Gordon, J. W., Scangos, G. A., Plotkin, D. J., Barbosa, J. A., and Ruddle, F. H. (1980) Genetic transformation of mouse embryos by microinjection of purified DNA. Proc. Natl. Acad. Sci. USA 77, 7380–7384.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (eds.) (1989) Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Schedl, A., Larin, Z., Montoliu, L., Thies, E., Kelsey, G., Lehrach, H., and Schütz, G. (1993) A method for the generation of YAC transgenic mice by pronuclear microinjection. Nucleic Acids Res. 21, 4783–4787.
Shizuya, H., Birren, B., Kim, U. J., Mancino, V., Slepak, T., Tachiiri, Y., and Simon, M. (1992) Cloning and stable maintainance of 300-kilobase-pair fragments of human DNA in Escherichia coli using an F-factor-based vector. Proc. Natl. Acad. Sci. USA 89, 8794–8797.
Ioannou, P. A., Amemiya, C. T., Garnes, J., Kroisel, P. M., Shizuya, H., Chen, C., Batzer, M. A., and de-Jong, P. J. (1994) A new bacteriophage P1-derived vector for the propagation of large human DNA fragments. Nat. Genet. 6, 84–89.
Lathe, R., Vilotte, J. L., and Clarke, A. J. (1987) Plasmid and bacteriophage vectors for the excision of intact inserts. Gene 57, 193–201.
Murphy, G. (1993) Generation of a nested set of deletions using exonuclease III, in Methods in Molecular Biology, vol. 23: DNA Sequencing Protocols (Griffin, H. G. and Griffin, A. M., eds.), Humana, Totowa, NJ, pp. 51–59.
Deng, W. P. and Nickoloff, J. A. (1992) Site-directed mutagenesis of virtually any plasmid by eliminating a unique site. Anal. Biochem. 200, 81–88.
Brinster, R. L., Allen, J. M., Behringer, R. R., Gelinas, R. E., and Palmiter, R. D. (1988) Introns increase transcriptional efficiency in transgenic mice. Proc. Natl. Acad. Sci. USA 85, 836–840.
Choi, T., Huang, M., Gorman, C., and Jaenisch, R. (1991) A generic intron increases gene expression in transgenic mice. Mol. Cell. Biol. 11, 3070–3074.
Lewis, B. A. and Orkin, S. H. (1995) A functional initiator element in the human β-globin promoter. J. Biol. Chem. 270, 28,139–28,144.
Gossler, A., Joyner, A. L., Rossant, J., and Skarnes, W. C. (1989) Mouse embryonic stem cells and reporter constructs to detect developmentally regulated genes. Science 244, 463–465.
Kothary, R., Clapoff, S., Darling, S., Perry, M. D., Moran, L. A., and Rossant, J. (1989) Inducible expression of an hsp68-lacZ hybrid gene in transgenic mice. Development 105, 707–714.
Gutman, A., Gilthorpe, J. D., and Rigby, P. W. J. (1994) Multiple positive and negative regulatory elements in the promoter of the mouse homeobox gene Hoxb-4. Mol. Cell. Biol. 14, 8143–8154.
Allen, N. D., Cran, D. G., Barton, S. C., Hettle, S., Reik, W., and Surani, M. A. (1988) Transgenes as probes for active chromosomal domains in mouse development. Nature 333, 852–855.
Goldhammer, D. J., Brunk, B. P., Faerman, A., King, A., Shani, M., and Emerson, C. P., Jr. (1995) Embryonic activation of the myoD gene is regulated by a highly conserved distal control element. Development 121, 637–649.
Luckow, B. and Schutz, G. (1987) CAT constructions with multiple unique restriction sites for the functional analysis of eukaryotic promoters and regulatory elements. Nucleic Acids Res. 15, 5490.
Schöler, H. R., Balling, R., Hatzopoulos, A. K., Suzuki, N., and Gruss, P. (1989) Octamer binding proteins confer transcriptional activity in early mouse embryogenesis. EMBO J. 8, 2551–2557.
Beddington, R. S. P., Morgernstern, J., Land, H., and Hogan, A. (1989) An in situ transgenic enzyme marker for the midgestation mouse embryo and the visualization of inner cell mass clones during early organogenesis. Development 106, 37–46.
Bronson, S. K., Plaehn, E. G., Kluckman, K. D., Hagaman, J. R., Maeda, N., and Smithies, O. (1996) Single-copy transgenic mice with chosen-site integration. Proc. Natl. Acad. Sci. USA 93, 9067–9072.
Gautier, C., Mehtali, M., and Lathe, R. (1989) A ubiquitous mammalian expression vector, pHMG, based on a housekeeping gene promoter. Nucleic Acids Res. 17, 8389.
Mehtali, M., LeMeur, M., and Lathe, R. (1990) The methylation-free status of a housekeeping transgene is lost at high copy number. Gene 91, 179–184.
Püschel, A., Balling, R., and Gruss, P. (1991) Separate elements cause lineage restriction and specify boundaries of Hox-1.1 expression. Development 112, 279–287.
Fire, A., Harrison, S. W., and Dixon, D. (1990) A modular set of lacZ fusion vectors for studying gene expression in Caenorhabditis elegans. Gene 93, 189–198.
Kam, W., Clauser, E., Kim, Y. S., Kan, Y. W., and Rutter, W. J. (1985) Cloning, sequencing and chromosomal localization of human term placental alkaline phosphatase cDNA. Proc. Natl. Acad. Sci. USA 82, 8715–8719.
Fields-Berry, S. C., Halliday, A. L., and Cepko, C. L. (1992) A recombinant retrovirus encoding alkaline phosphatase confirms clonal boundary assignment in lineage analysis of murine retina. Proc. Natl. Acad. Sci. USA 89, 693–697.
Chiu, M. I. and Nathans, J. (1994) Blue cones and cone bipolar cells share transcriptional specificity as determined by expression of human blue pigment-derived transgenes. J. Neurosci. 14, 3426–3436.
Chalfie, M., Tu, Y., Euskirchen, G., Ward, W. W., and Prasher, D. C. (1994) Green fluorescent protein as a marker for gene expression. Science 263, 802–805.
Kain, S. R., Adams, M., Kondepudi, A., Yang, T. T., Ward, W. W., and Kitts, P. (1995) Green fluorescent protein as a reporter of gene expression and protein localization. Biotechniques 19, 650–655.
De Giorgi, F., Brini, M., Bastianutto, C., Marsault, R., Montero, M., Pizzo, P., et al. (1996) Targeting aequorin and green fluorescent protein to intracellular organelles. Gene 173, 113–117.
Plautz, J. D., Day, R. N., Dailey, G. M., Welsh, S. B., Hall, J. C., Halpain, S., et al. (1996) Green fluorescent protein and its derivatives as versatile markers for gene expression in living Drosophila melanogaster, plant and mammalian cells. Gene 173, 83–87.
Amsterdam, A., Lin, S., Moss, L. G., and Hopkins, N. (1996) Requirements for green fluorescent protein detection in transgenic zebrafish embryos. Gene 173, 99–103.
Ogawa, H., Inouye, S., Tsuji, F. I., Yasuda, K., and Umesono, K. (1995) Localization, trafficking, and temperature-dependence of the Aequorea green fluorescent protein in cultured vertebrate cells. Proc. Natl. Acad. Sci. USA 92, 11,899–11,903.
Heim, R., Prasher, D. C., and Tsien, R. Y. (1994) Wavelength mutations and posttranslational autoxidation of green fluorescent protein. Proc. Natl. Acad. Sci. USA 91, 12,501–12,504.
Delagrave, S., Hawtin, R. E., Silva, C. M., Yang, M. M., and Youvan, D. C. (1995) Redshifted excitation mutants of the green fluorescent protein. Biotechnology 13, 151–154.
Ehrig, T., O’Kane, D. J., and Prendergast, F. G. (1995) Green fluorescent protein mutants with altered fluorescence excitation spectra. FEBS Lett. 367, 163–166.
Heim, R., Cubitt, A. B., and Tsien, R. Y. (1995) Improved green fluorescence. Nature 373, 663,664.
Cormack, B. P., Valdivia, R. H., and Falkow, S. (1996) FACS-optimized mutants of the green fluorescent protein (GFP). Gene 173, 33–38.
Heim, R. and Tsien, R. Y. (1996) Engineering green fluorescent protein for improved brightness, longer wavelengths and fluorescence resonance energy transfer. Curr. Biol. 6, 178–182.
Anderson, M. T., Tjioe, I. M., Lorincz, M. C., Parks, D. R., Herzenberg, L. A., Nolan, G. P., and Herzenberg, L. A. (1996) Simultaneous fluorescent-activated cell sorter analysis of two distinct transcriptional elements within a single cell using engineered green fluorescent proteins. Proc. Natl. Acad. Sci. USA 93, 8508–8511.
Zolotukhin, S., Potter, M., Hauswirth, W. W., Guy, J., and Muzyczka, N. (1996) A “humanized” green fluorescent protein cDNA adapted for high-level expression in mammalian cells. J. Virol. 70, 4646–4654.
Aguzzi, A. and Theuring, F. (1994) Improved in situ beta-galactosidase staining for histological analysis of transgenic mice. Histochemistry 102, 477–481.
Callahan, C. A. and Thomas, J. B. (1994) Tau-β-galactosidase, an axon-targetted fusion protein. Proc. Natl. Acad. Sci. USA 91, 5972–5976.
Gorman, C., Padmanabhan, R., and Howard, B. H. (1983) High efficiency DNA-mediated transformation of primate cells. Science 221, 551–553.
Brinster, R. L., Chen, H. Y., Trumbauer, M. E., Yagle, M. K., and Palmiter, R. D. (1985) Factors affecting the efficiency of introducing foreign DNA into mice by microinjecting eggs. Proc. Natl. Acad. Sci. USA 82, 4438–4442.
Townes, T. M., Lingrel, J. B., Chen, H. Y., Brinster, R. L., and Palmiter, R. D. (1985) Erythroid specific expression of human beta-globin genes in transgenic mice. EMBO J. 4, 1715–1723.
Jackson, R. J., Howell, M. T., and Kaminski, A. (1990) The novel mechanism of initiation of picornavirus RNA translation. Trends Biochem. Sci. 15, 477–483.
Kozak, M. (1989) The scanning model for translation: an update. J. Cell Biol. 108, 229–241.
Kim, D. G., Kang, H. M., Jang, S. K., and Shin, H. S. (1992) Construction of a bifunctional mRNA in the mouse by using the internal ribosome entry site of encephalomyocarditis virus. Mol. Cell. Biol. 12, 3636–3643.
Curatola, A. M., Nadal, M. S., and Schneider, R. J. (1995) Rapid degradation of AU-rich element (ARE) mRNAs is activated by ribosome transit and blocked by secondary structure at any position 5′ to the ARE. Mol. Cell. Biol. 15, 6331–6340.
Sachs, A. B. (1993) Messenger RNA degradation in eukaryotes. Cell 74, 413–421.
Hahnel, A. C., Rappolee, D. A., Millan, J. L., Manes, T., Ziomek, C. A., Theodosiou, N. G., Werb, Z., Pedersen, R. A., and Schultz, G. A. (1990) Two alkaline phosphatase genes are expressed during early development in the mouse embryo. Development 110, 555–564.
Wingender, E. (1994) Recognition of regulatory regions in genomic sequences. J. Biotechnol. 35, 273–280.
Gundersen, K., Sanes, J. R., and Merlie, J. P. (1993) Neural regulation of muscle acetylcholine receptor epsilon-and alpha-subunit gene promoters in transgenic mice. J. Cell Biol. 123, 1535–1544.
Echelard, Y., Vassileva, G., and McMahon, A. P. (1994) Cis-acting regulatory sequences governing Wnt-1 expression in the developing mouse CNS. Development 120, 2213–2224.
Gérard, M., Duboule, D., and Zákány, J. (1993) Structure and activity of regulatory elements involved in the activation of the Hoxd-11 gene during late gastrulation. EMBO J. 12, 3539–3550.
Logan, C., Khoo, W. K., Cado, D., and Joyner, A. L. (1993) Two enhancer regions in the mouse En-2 locus direct expression to the mid/hindbrain region and mandibular myoblasts. Development 117, 905–916.
Song, D.-L, Chalepakis, G., Gruss, P., and Joyner, A. L. (1996) Two Pax-binding sites are required for early embryonic brain expression of an Engrailed-2 transgene. Development 122, 627–635.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1999 Humana Press Inc.
About this protocol
Cite this protocol
Gilthorpe, J.D., Rigby, P.W.J. (1999). Reporter Genes for the Study of Transcriptional Regulation in Transgenic Mouse Embryos. In: Sharpe, P.T., Mason, I. (eds) Molecular Embryology. Methods in Molecular Biology™, vol 97. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-270-8:159
Download citation
DOI: https://doi.org/10.1385/1-59259-270-8:159
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-0-89603-387-0
Online ISBN: 978-1-59259-270-8
eBook Packages: Springer Protocols