Abstract
Transcriptional feedback loops are at the core of the molecular clockworks. As single clock genes were cloned it was compelling to develop an assay that allowed simple and direct functional testing of putative activators or repressors of transcription. This chapter includes a general description and guidelines to carry out transcriptional assays in transiently transfected Schneider’s cells.
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References
Darlington, T. K., Wager-Smith, K., Ceriani, M. F., et al. (1998) Closing the circadian loop: CLOCK-induced transcription of its own inhibitors per and tim. Science 280, 1599–1603.
Froy, O., Chang, D.C., and Reppert, S.M. (2002) Redox potential: differential roles in dCRY and mCRY1 functions. Curr. Biol. 12, 147–152.
Chang, D. C., McWatters, H. G., Williams, J. A., Gotter, A. L., Levine, J. D., and Reppert, S. M. (2003) Constructing a feedback loop with circadian clock molecules from the silkmoth, Antheraea pernyi. J. Biol. Chem. 278, 38,149–38,158.
Ceriani, M. F., Darlington, T. K., Staknis, D., et al. (1999). Light-dependent sequestration of TIMELESS by CRYPTOCHROME. Science 285, 553–556.
Ko, H. W., Jiang, J., and Edery, I. (2002) Role for Slimb in the degradation of Drosophila Period protein phosphorylated by Doubletime. Nature 420, 673–678.
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.
von Ohlen, T., Lessing, D., Nusse, R., and Hooper, J. E. (1997) Hedgehog signaling regulates transcription through cubitus interruptus, a sequence-specific DNA binding protein. Proc. Natl. Acad. Sci. USA 94, 2404–2409.
Brandes, C., Plautz, J. D., Stanewsky, R., et al. (1996) Novel features of Drosophila period transcription revealed by real-time luciferase reporting. Neuron 16, 687–692.
Plautz, J. D., Straume, M., Stanewsky, R., et al. (1997) Quantitative analysis of Drosophila period gene transcription in living animals. J. Biol. Rhythms 12, 204–217.
Stanewsky, R., Jamison, C. F., Plautz, J. D., Kay, S. A., and Hall, J. C. (1997) Multiple circadian-regulated elements contribute to cycling period gene expression in Drosophila. EMBO J. 16, 5006–5018.
Plautz, J. D., Kaneko, M., Hall, J.C., and Kay, S. A. (1997) Independent photoreceptive circadian clocks throughout Drosophila. Science 278, 1632–1635.
Hao, H., Allen, D. L. and Hardin, P. E. (1997) A circadian enhancer mediates PER-dependent mRNA cycling in Drosophila melanogaster. Mol. Cell Biol. 17, 3687–3693.
Froy, O., Chang, D.C., and Reppert, S. M. (2002) Redox potential: differential roles in dCRY and mCRY1 functions. Curr. Biol. 12, 147–152.
Sonnenfeld, M., Ward, M., Nystrom, G., Mosher, J., Stahl, S., and Crews, S. (1997) The Drosophila tango gene encodes a bHLH-PAS protein that is orthologous to mammalian Arnt and controls CNS midline and tracheal development. Development 124, 4571–4582.
Nawathean, P., and Rosbash, M. (2004) The doubletime and CKII kinases collaborate to potentiate Drosophila PER transcriptional repressor activity. Mol. Cell 13, 213–223.
Weber, F., and Kay, S. A. (2003) A PERIOD inhibitor buffer introduces a delay mechanism for CLK/CYC-activated transcription. FEBS Lett. 555, 341–345.
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© 2007 Humana Press Inc.
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Ceriani, M.F. (2007). Reporter Assays. In: Rosato, E. (eds) Circadian Rhythms. Methods in Molecular Biology™, vol 362. Humana Press. https://doi.org/10.1007/978-1-59745-257-1_37
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DOI: https://doi.org/10.1007/978-1-59745-257-1_37
Publisher Name: Humana Press
Print ISBN: 978-1-58829-417-3
Online ISBN: 978-1-59745-257-1
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