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Related Experiment Video

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Immunostaining for DNA Modifications: Computational Analysis of Confocal Images
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Imaging Histone Methylations in Living Animals.

Thillai V Sekar1, Ramasamy Paulmurugan2,3

  • 1Molecular Imaging Program at Stanford, Bio-X Program, Stanford University School of Medicine, Stanford, CA, 94305, USA.

Methods in Molecular Biology (Clifton, N.J.)
|July 19, 2016
PubMed
Summary

Researchers developed novel optical imaging sensors to monitor histone methylation, a key regulator of gene expression. These tools enable drug screening and preclinical evaluation for diseases linked to histone methylation dysregulation.

Keywords:
DegronHistone methylationIn vivo imagingLuciferaseOptical imagingProteaseReporter genesSplit reporters

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Area of Science:

  • Molecular Biology
  • Epigenetics
  • Biochemistry

Background:

  • Histone modifications regulate gene expression and cellular networks.
  • Histone lysine methylation is crucial for gene regulation in heterochromatic regions.
  • Dysregulated histone methylation is implicated in various diseases, making it a therapeutic target.

Purpose of the Study:

  • To develop novel methods for screening and preclinical evaluation of drugs targeting histone methylation.
  • To create optical imaging sensors for monitoring histone methylation levels in cells and living animals.

Main Methods:

  • Development of two independent optical imaging sensors utilizing reporter gene technology.
  • Application of standard PCR-based cloning strategies for plasmid vector construction.
  • Detailed methods for measuring histone methylation-assisted luciferase quantitation in cells and imaging in living animals.

Main Results:

  • Successful development of two distinct optical imaging sensors for histone methylation.
  • Demonstration of quantitative monitoring of histone methylation levels in intact cells.
  • Capability to image histone methylation in living animals for preclinical drug evaluation.

Conclusions:

  • The developed optical imaging sensors provide a simple method for screening drugs that modulate histone lysine methylation.
  • These tools are valuable for preclinical evaluation of potential therapeutics targeting histone methylation.
  • The sensors facilitate a deeper understanding of histone methylation's role in cellular processes and disease.