Inducible expression of an hsp68-lacZ hybrid gene in transgenic mice

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Inducible expression of an hsp68-lacZ hybrid gene in transgenic mice

R Kothary, S Clapoff, S Darling, MD Perry, LA Moran and J Rossant
Division of Molecular and Developmental Biology, Mt. Sinai Hospital Research Institute, Toronto, Ontario, Canada.

Transgenic mice have been generated that express the E. coli beta-galactosidase gene under the control of the promoter from the mouse heat-shock gene, hsp68. Sequences from -664 to +113 relative to the start of transcription of the hsp68 gene were sufficient to direct stress-induced expression of the beta-galactosidase gene in adult tail tissue and various tissues of fetal stages of development. Expression was detected in situ by staining with the chromogenic substrate, X-gal. The hybrid gene was refractory to induction in preimplantation embryos until the blastocyst stage of development, as reported for the endogenous hsp68 gene. No constitutive expression was observed by in situ staining or Northern analysis at any stage of development, even in tissues that constitutively express the endogenous hsp68 gene. We conclude that the hsp68 promoter region included in the construct contains sufficient sequence information for heat and arsenite inducibility, but it does not contain sequences controlling tissue-specific expression during development. This tightly regulated inducible promoter may provide a useful tool for short-term inducible gene expression in transgenic mice.

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				H. Wu, S.-c. Kao, T. Barrientos, S. H. Baldwin, E. N. Olson, G. R. Crabtree, B. Zhou, and C.-P. Chang Down Syndrome Critical Region-1 Is a Transcriptional Target of Nuclear Factor of Activated T Cells-c1 within the Endocardium during Heart Development J. Biol. Chem., October 19, 2007; 282(42): 30673 - 30679. [Abstract] [Full Text] [PDF]

				T. Werner, A. Hammer, M. Wahlbuhl, M. R. Bosl, and M. Wegner Multiple conserved regulatory elements with overlapping functions determine Sox10 expression in mouse embryogenesis Nucleic Acids Res., October 8, 2007; 35(19): 6526 - 6538. [Abstract] [Full Text] [PDF]

				A. Visel, S. Minovitsky, I. Dubchak, and L. A. Pennacchio VISTA Enhancer Browser--a database of tissue-specific human enhancers Nucleic Acids Res., January 12, 2007; 35(suppl_1): D88 - D92. [Abstract] [Full Text] [PDF]

				S. Ribich, B. Tasic, and T. Maniatis Identification of long-range regulatory elements in the protocadherin-{alpha} gene cluster PNAS, December 26, 2006; 103(52): 19719 - 19724. [Abstract] [Full Text] [PDF]

				S. Prabhakar, F. Poulin, M. Shoukry, V. Afzal, E. M. Rubin, O. Couronne, and L. A. Pennacchio Close sequence comparisons are sufficient to identify human cis-regulatory elements Genome Res., July 1, 2006; 16(7): 855 - 863. [Abstract] [Full Text] [PDF]

				Y. Nakajima, M. Morimoto, Y. Takahashi, H. Koseki, and Y. Saga Identification of Epha4 enhancer required for segmental expression and the regulation by Mesp2 Development, July 1, 2006; 133(13): 2517 - 2525. [Abstract] [Full Text] [PDF]

				M. Zubair, S. Ishihara, S. Oka, K. Okumura, and K.-i. Morohashi Two-Step Regulation of Ad4BP/SF-1 Gene Transcription during Fetal Adrenal Development: Initiation by a Hox-Pbx1-Prep1 Complex and Maintenance via Autoregulation by Ad4BP/SF-1. Mol. Cell. Biol., June 1, 2006; 26(11): 4111 - 4121. [Abstract] [Full Text] [PDF]

				X. Chi, P. K. Chatterjee, W. Wilson III, S.-X. Zhang, F. J. DeMayo, and R. J. Schwartz Complex cardiac Nkx2-5 gene expression activated by noggin-sensitive enhancers followed by chamber-specific modules PNAS, September 20, 2005; 102(38): 13490 - 13495. [Abstract] [Full Text] [PDF]

				A. Rojas, S. De Val, A. B. Heidt, S.-M. Xu, J. Bristow, and B. L. Black Gata4 expression in lateral mesoderm is downstream of BMP4 and is activated directly by Forkhead and GATA transcription factors through a distal enhancer element Development, August 1, 2005; 132(15): 3405 - 3417. [Abstract] [Full Text] [PDF]

				J. P. Konhilas, U. Widegren, D. L. Allen, A. C. Paul, A. Cleary, and L. A. Leinwand Loaded wheel running and muscle adaptation in the mouse Am J Physiol Heart Circ Physiol, July 1, 2005; 289(1): H455 - H465. [Abstract] [Full Text] [PDF]

				D. Phan, T. L. Rasmussen, O. Nakagawa, J. McAnally, P. D. Gottlieb, P. W. Tucker, J. A. Richardson, R. Bassel-Duby, and E. N. Olson BOP, a regulator of right ventricular heart development, is a direct transcriptional target of MEF2C in the developing heart Development, June 1, 2005; 132(11): 2669 - 2678. [Abstract] [Full Text] [PDF]

				J. K. Takeuchi, M. Mileikovskaia, K. Koshiba-Takeuchi, A. B. Heidt, A. D. Mori, E. P. Arruda, M. Gertsenstein, R. Georges, L. Davidson, R. Mo, et al. Tbx20 dose-dependently regulates transcription factor networks required for mouse heart and motoneuron development Development, May 15, 2005; 132(10): 2463 - 2474. [Abstract] [Full Text] [PDF]

				J. Y. H. Li, Z. Lao, and A. L. Joyner New regulatory interactions and cellular responses in the isthmic organizer region revealed by altering Gbx2 expression Development, April 15, 2005; 132(8): 1971 - 1981. [Abstract] [Full Text] [PDF]

				F. J. Davis, J. B Pillai, M. Gupta, and M. P. Gupta Concurrent opposite effects of trichostatin A, an inhibitor of histone deacetylases, on expression of {alpha}-MHC and cardiac tubulins: implication for gain in cardiac muscle contractility Am J Physiol Heart Circ Physiol, March 1, 2005; 288(3): H1477 - H1490. [Abstract] [Full Text] [PDF]

				J. P. Konhilas, A. H. Maass, S. W. Luckey, B. L. Stauffer, E. N. Olson, and L. A. Leinwand Sex modifies exercise and cardiac adaptation in mice Am J Physiol Heart Circ Physiol, December 1, 2004; 287(6): H2768 - H2776. [Abstract] [Full Text] [PDF]

				J. Galceran, C. Sustmann, S.-C. Hsu, S. Folberth, and R. Grosschedl LEF1-mediated regulation of Delta-like1 links Wnt and Notch signaling in somitogenesis Genes & Dev., November 15, 2004; 18(22): 2718 - 2723. [Abstract] [Full Text] [PDF]

				T. Hu, H. Yamagishi, J. Maeda, J. McAnally, C. Yamagishi, and D. Srivastava Tbx1 regulates fibroblast growth factors in the anterior heart field through a reinforcing autoregulatory loop involving forkhead transcription factors Development, November 1, 2004; 131(21): 5491 - 5502. [Abstract] [Full Text] [PDF]

				S. M. Brugger, A. E. Merrill, J. Torres-Vazquez, N. Wu, M.-C. Ting, J. Y.-M. Cho, S. L. Dobias, S. E. Yi, K. Lyons, J. R. Bell, et al. A phylogenetically conserved cis-regulatory module in the Msx2 promoter is sufficient for BMP-dependent transcription in murine and Drosophila embryos Development, October 15, 2004; 131(20): 5153 - 5165. [Abstract] [Full Text] [PDF]

				K.-H. Lee, S. Evans, T. Y. Ruan, and A. B. Lassar SMAD-mediated modulation of YY1 activity regulates the BMP response and cardiac-specific expression of a GATA4/5/6-dependent chick Nkx2.5 enhancer Development, October 1, 2004; 131(19): 4709 - 4723. [Abstract] [Full Text] [PDF]

				E. Dodou, M. P. Verzi, J. P. Anderson, S.-M. Xu, and B. L. Black Mef2c is a direct transcriptional target of ISL1 and GATA factors in the anterior heart field during mouse embryonic development Development, August 15, 2004; 131(16): 3931 - 3942. [Abstract] [Full Text] [PDF]

				J. P. Anderson, E. Dodou, A. B. Heidt, S. J. De Val, E. J. Jaehnig, S. B. Greene, E. N. Olson, and B. L. Black HRC Is a Direct Transcriptional Target of MEF2 during Cardiac, Skeletal, and Arterial Smooth Muscle Development In Vivo Mol. Cell. Biol., May 1, 2004; 24(9): 3757 - 3768. [Abstract] [Full Text] [PDF]

				C. Kimura-Yoshida, K. Kitajima, I. Oda-Ishii, E Tian, M. Suzuki, M. Yamamoto, T. Suzuki, M. Kobayashi, S. Aizawa, and I. Matsuo Characterization of the pufferfish Otx2 cis-regulators reveals evolutionarily conserved genetic mechanisms for vertebrate head specification Development, January 1, 2004; 131(1): 57 - 71. [Abstract] [Full Text] [PDF]

				F. Santagati, K. Abe, V. Schmidt, T. Schmitt-John, M. Suzuki, K.-i. Yamamura, and K. Imai Identification of Cis-regulatory Elements in the Mouse Pax9/Nkx2-9 Genomic Region: Implication for Evolutionary Conserved Synteny Genetics, September 1, 2003; 165(1): 235 - 242. [Abstract] [Full Text] [PDF]

				L. T. Krebs, N. Iwai, S. Nonaka, I. C. Welsh, Y. Lan, R. Jiang, Y. Saijoh, T. P. O'Brien, H. Hamada, and T. Gridley Notch signaling regulates left-right asymmetry determination by inducing Nodal expression Genes & Dev., May 15, 2003; 17(10): 1207 - 1212. [Abstract] [Full Text] [PDF]

				S. J. Kwang, S. M. Brugger, A. Lazik, A. E. Merrill, L.-Y. Wu, Y.-H. Liu, M. Ishii, F. O. Sangiorgi, M. Rauchman, H. M. Sucov, et al. Msx2 is an immediate downstream effector of Pax3 in the development of the murine cardiac neural crest Development, March 3, 2003; 129(2): 527 - 538. [Abstract] [Full Text] [PDF]

				H. Yamagishi, J. Maeda, T. Hu, J. McAnally, S. J. Conway, T. Kume, E. N. Meyers, C. Yamagishi, and D. Srivastava Tbx1 is regulated by tissue-specific forkhead proteins through a common Sonic hedgehog-responsive enhancer Genes & Dev., January 15, 2003; 17(2): 269 - 281. [Abstract] [Full Text] [PDF]

				L. Huang, N. F. Mivechi, and D. Moskophidis Insights into Regulation and Function of the Major Stress-Induced hsp70 Molecular Chaperone In Vivo: Analysis of Mice with Targeted Gene Disruption of the hsp70.1 or hsp70.3 Gene Mol. Cell. Biol., December 15, 2001; 21(24): 8575 - 8591. [Abstract] [Full Text] [PDF]