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Author Spotlight: Enhancements in Gene Expression Regulation Research
Published on: September 15, 2023
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Quantification of Global Histone Post Translational Modifications Using Intranuclear Flow Cytometry in Isolated Mouse
Morgan Towriss1, Jennifer Kim2, Annie Vogel Ciernia3
1Department of Biochemistry and Molecular Biology, Djavad Mowafaghian Centre for Brain Health, University of British Columbia.
Journal of Visualized Experiments : Jove
|October 2, 2023
Summary
This study introduces a new flow cytometry method to rapidly measure global histone modifications. This technique allows for high-throughput screening of epigenetic changes in cells and tissues.
Area of Science:
- Epigenetics
- Molecular Biology
- Cell Biology
Background:
- Gene expression is regulated by chromatin structure and histone post-translational modifications (HPTMs).
- HPTMs dynamically influence gene expression, acting as repressors or activators.
- Current methods, like next-generation sequencing, analyze only one HPTM at a time, limiting comprehensive analysis.
Purpose of the Study:
- To develop a high-throughput, quantitative method for assessing global HPTMs.
- To enable screening of histone modifications before large-scale genomic studies.
- To provide a versatile protocol applicable to various cellular contexts.
Main Methods:
- A flow cytometry-based protocol was developed to detect global HPTMs.
- The method can be applied to cells in culture or isolated cells from tissues.
- Example data demonstrated sensitivity using mouse brain microglia stimulated with lipopolysaccharide.
Main Results:
- The developed protocol provides a rapid and quantitative measure of global HPTMs.
- The assay successfully detected global HPTM shifts in response to immune stimuli.
- The method is sensitive and applicable to diverse biological samples.
Conclusions:
- This flow cytometry protocol offers a powerful tool for high-throughput screening of global HPTMs.
- The method facilitates the study of epigenetic regulation in various cellular states and stimuli.
- It can be adapted to analyze any transcriptional or epigenetic regulator detectable by antibodies.
