Trinucleotide Repeat Protocols pp 261-276

Part of the Methods in Molecular Biology™ book series (MIMB, volume 277)

Chromatin Immunoprecipitation Technique for Study of Transcriptional Dysregulation in Intact Mouse Brain

  • Melissa W. Braveman
  • Alice S. Chen-Plotkin
  • George J. Yohrling
  • Jang-Ho J. Cha

Summary

Transcriptional dysregulation has emerged as an important pathologic mechanism underlying the pathogenesis of Huntington’s disease (HD). The control of transcription depends on appropriate binding of transcription factor proteins to specific promoter regions of genes. Chromatin immunoprecipitation (ChIP) is a technique that has been used to study the association of transcription factors with DNA. To address the hypothesis that there is altered transcription factor-DNA association in HD, we have recently adapted the ChIP technique to the study of transgenic mouse brain. Here, we describe our method of performing ChIP in intact mouse brain. We have optimized conditions for formaldehyde crosslinking, antibody immunoprecipitation, and quantitative real-time polymerase chain reaction detection. Using ChIP, one can measure the association of transcription factors with specific genes and determine if this association is altered in transgenic HD mouse models. ChIP applied to whole-mouse brain can thus offer a window into mechanisms of transcriptional dysregulation.

Key Words

Transcription transcription factor mRNA DNA histone Sp1 real-time PCR immunoprecipitation chromatin 

References

  1. 1.
    Cha, J.-H. J. (2000) Transcriptional dysregulation in Huntington’s disease. Trends Neurosci. 23, 387–392.PubMedCrossRefGoogle Scholar
  2. 2.
    Cha, J.-H. J., Kosinski, C. M., Kerner, J. A., et al. (1998) Altered brain neurotransmitter receptors in transgenic mice expressing a portion of an abnormal human Huntington disease gene. Proc. Natl. Acad. Sci. USA 95, 6480–6485.PubMedCrossRefGoogle Scholar
  3. 3.
    Cha, J.-H. J., Frey, A. S., Alsdorf, S. A., et al. (1999) Altered neurotransmitter receptor expression in transgenic mouse models of Huntington’s disease. Phil. Trans. R. Soc. London B: Biol. Sci. 354, 981–989.CrossRefGoogle Scholar
  4. 4.
    Luthi-Carter, R., Strand, A., Peters, N. L., et al. (2000) Decreased expression of striatal signaling genes in a mouse model of Huntington’s disease. Hum. Mol. Genet. 9, 1259–1271.PubMedCrossRefGoogle Scholar
  5. 5.
    Shang, Y., Hu, X., DiRenzo, J., et al. (2000) Cofactor dynamics and sufficiency in estrogen receptor-regulated transcription. Cell 103, 843–852.PubMedCrossRefGoogle Scholar
  6. 6.
    Noma, K., Allis, C. D., and Grewal, S. I. (2001) Transitions in distinct histone H3 methylation patterns at the heterochromatin domain boundaries. Science 293, 1150–1155.PubMedCrossRefGoogle Scholar
  7. 7.
    Lo, W. S., Duggan, L., Tolga, N. C., et al. (2001) Snf1—a histone kinase that works in concert with the histone acetyltransferase Gcn5 to regulate transcription. Science 293, 1142–1146.PubMedCrossRefGoogle Scholar
  8. 8.
    Breiling, A., Turner, B. M., Bianchi, M. E., et al. (2001) General transcription factors bind promoters repressed by Polycomb group proteins. Nature 412, 651–655.PubMedCrossRefGoogle Scholar
  9. 9.
    Chen, H., Lin, R. J., Xie, W., et al. (1999) Regulation of hormone-induced histone hyperacetylation and gene activation via acetylation of an acetylase. Cell 98, 675–686.PubMedCrossRefGoogle Scholar
  10. 10.
    Gerhold, D., Rushmore, T., and Caskey, C. T. (1999) DNA chips: promising toys have become powerful tools. Trends Biochem. Sci. 24, 168–173.PubMedCrossRefGoogle Scholar
  11. 11.
    Luo, R. X., Postigo, A. A., and Dean, D. C. (1998) Rb interacts with histone deacetylase to repress transcription. Cell 92, 463–473.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 2004

Authors and Affiliations

  • Melissa W. Braveman
    • 1
  • Alice S. Chen-Plotkin
    • 1
  • George J. Yohrling
    • 1
  • Jang-Ho J. Cha
    • 1
  1. 1.Mass General Institute for Neurodegenerative Disease and Department of NeurologyMassachusetts General HospitalCharlestown

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