Inducible and reversible Clock gene expression in brain using the tTA system for the study of circadian behavior.

Hee Kyung Hong, Jason L. Chong, Weimin Song, Eun Joo Song, Amira A. Jyawook, Andrew C. Schook, Caroline H. Ko, Joseph S. Takahashi

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

The mechanism of circadian oscillations in mammals is cell autonomous and is generated by a set of genes that form a transcriptional autoregulatory feedback loop. While these "clock genes" are well conserved among animals, their specific functions remain to be fully understood and their roles in central versus peripheral circadian oscillators remain to be defined. We utilized the in vivo inducible tetracycline-controlled transactivator (tTA) system to regulate Clock gene expression conditionally in a tissue-specific and temporally controlled manner. Through the use of Secretogranin II to drive tTA expression, suprachiasmatic nucleus- and brain-directed expression of a tetO::Clock(Delta19) dominant-negative transgene lengthened the period of circadian locomotor rhythms in mice, whereas overexpression of a tetO::Clock(wt) wild-type transgene shortened the period. Low doses (10 mug/ml) of doxycycline (Dox) in the drinking water efficiently inactivated the tTA protein to silence the tetO transgenes and caused the circadian periodicity to return to a wild-type state. Importantly, low, but not high, doses of Dox were completely reversible and led to a rapid reactivation of the tetO transgenes. The rapid time course of tTA-regulated transgene expression demonstrates that the CLOCK protein is an excellent indicator for the kinetics of Dox-dependent induction/repression in the brain. Interestingly, the daily readout of circadian period in this system provides a real-time readout of the tTA transactivation state in vivo. In summary, the tTA system can manipulate circadian clock gene expression in a tissue-specific, conditional, and reversible manner in the central nervous system. The specific methods developed here should have general applicability for the study of brain and behavior in the mouse.

Original languageEnglish (US)
JournalPLoS Genetics
Volume3
Issue number2
DOIs
StatePublished - Feb 23 2007

Fingerprint

Trans-Activators
gene expression
brain
Transgenes
transgenes
Gene Expression
doxycycline
Doxycycline
Brain
circadian rhythm
protein
gene
nervous system
reactivation
CLOCK Proteins
periodicity
Secretogranin II
mammal
oscillation
drinking water

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology
  • Ecology, Evolution, Behavior and Systematics
  • Cancer Research
  • Genetics(clinical)

Cite this

Inducible and reversible Clock gene expression in brain using the tTA system for the study of circadian behavior. / Hong, Hee Kyung; Chong, Jason L.; Song, Weimin; Song, Eun Joo; Jyawook, Amira A.; Schook, Andrew C.; Ko, Caroline H.; Takahashi, Joseph S.

In: PLoS Genetics, Vol. 3, No. 2, 23.02.2007.

Research output: Contribution to journalArticle

Hong, Hee Kyung ; Chong, Jason L. ; Song, Weimin ; Song, Eun Joo ; Jyawook, Amira A. ; Schook, Andrew C. ; Ko, Caroline H. ; Takahashi, Joseph S. / Inducible and reversible Clock gene expression in brain using the tTA system for the study of circadian behavior. In: PLoS Genetics. 2007 ; Vol. 3, No. 2.
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