A CRISPR-based approach for proteomic analysis of a single genomic locus

Zachary J. Waldrip, Stephanie D. Byrum, Aaron J. Storey, Jun Gao, Alicia K. Byrd, Samuel G. Mackintosh, Wayne P. Wahls, Sean D. Taverna, Kevin D. Raney, Alan J. Tackett

Research output: Contribution to journalArticle

Abstract

Any given chromosomal activity (e.g., transcription) is governed predominantly by the local epiproteome. However, defining local epiproteomes has been limited by a lack of effective technologies to isolate discrete sections of chromatin and to identify with precision specific proteins and histone posttranslational modifications (PT Ms). We report the use of the Cas9 and guide RNA (gRNA) components of the CRI SPR system for gRNA-directed purification of a discrete section of chromatin. Quantitative mass spectrometry provides for unambiguous identification of proteins and histone PT Ms specifically associated with the enriched chromatin. This CRI SPR -based Chromatin Affinity Purification with Mass Spectrometry (CRI SPR -ChAP-MS) approach revealed changes in the local epiproteome of a promoter during activation of transcription. CRI SPR -ChAP-MS thus has broad applications for discovering molecular components and dynamic regulation of any in vivo activity at a given chromosomal location.

Original languageEnglish (US)
Pages (from-to)1207-1211
Number of pages5
JournalEpigenetics
Volume9
Issue number9
DOIs
StatePublished - Jan 1 2014

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Keywords

  • Affinity purification
  • Epigenetics
  • Epiproteome
  • Histone
  • Posttranslational modification
  • Proteomics

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research

Cite this

Waldrip, Z. J., Byrum, S. D., Storey, A. J., Gao, J., Byrd, A. K., Mackintosh, S. G., Wahls, W. P., Taverna, S. D., Raney, K. D., & Tackett, A. J. (2014). A CRISPR-based approach for proteomic analysis of a single genomic locus. Epigenetics, 9(9), 1207-1211. https://doi.org/10.4161/epi.29919