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Related Experiment Videos

ChIP-re-ChIP: Co-occupancy Analysis by Sequential Chromatin Immunoprecipitation.

Timothy V Beischlag1, Gratien G Prefontaine2, Oliver Hankinson3

  • 1Faculty of Health Sciences, Simon Fraser University, 8888 University Drive, Burnaby, BC, Canada, V5A 1S6. tvb@sfu.ca.

Methods in Molecular Biology (Clifton, N.J.)
|October 14, 2017
PubMed
Summary

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Sequential ChIP (Chromatin Immunoprecipitation) assays identify protein complexes regulating gene transcription. This method enhances the analysis of transcriptional regulators and epigenetic modifications for greater definition.

Area of Science:

  • Molecular Biology
  • Genetics
  • Epigenetics

Background:

  • Chromatin immunoprecipitation (ChIP) analyzes DNA-associated protein interactions.
  • Identifying multi-protein complexes is crucial for understanding gene regulation.
  • Sequential ChIP (ChIP-re-ChIP) allows for deeper analysis of protein interactions at specific genomic locations.

Purpose of the Study:

  • To describe a simple and reliable method for performing sequential ChIP assays.
  • To enhance the power and definition of analyses for transcriptional regulators and cistromes.

Main Methods:

  • Utilizing the specific interactions between DNA and DNA-associated proteins.
  • Performing sequential ChIP (ChIP-re-ChIP) to examine multiple proteins at the same genomic site.
Keywords:
ChIP-re-ChIPChemical cross-linkingChromatin immunoprecipitation (ChIP)Co-occupancyPolymerase chain reaction (PCR)Promoter/regulatory region occupancyProtein–protein interactions

Related Experiment Videos

Main Results:

  • The described method allows for the identification of multi-protein chromatin complexes.
  • Enables the examination of specific transcriptional or epigenetic regulators.
  • Facilitates the identification of cistromes with higher precision.

Conclusions:

  • Sequential ChIP is a powerful technique for dissecting complex regulatory mechanisms.
  • The presented method offers a reliable approach for advanced chromatin analysis.