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

作者：Waldrip Zachary J; Byrum Stephanie D; Storey Aaron J; Gao Jun; Byrd Alicia K; Mackintosh Samuel G; Wahls Wayne P; Taverna Sean D; Raney Kevin D; Tackett Alan J^*

来源：Epigenetics, 2014, 9(9): 1207-1211.

DOI：10.4161/epi.29919

• epigenetics
• proteomics
• histone
• epiproteome
• posttranslational modification
• affinity purification

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 (PTMs). We report the use of the Cas9 and guide RNA (gRNA) components of the CRISPR system for gRNA-directed purification of a discrete section of chromatin. Quantitative mass spectrometry provides for unambiguous identification of proteins and histone PTMs specifically associated with the enriched chromatin. This CRISPR-based Chromatin Affinity Purification with Mass Spectrometry (CRISPR-ChAP-MS) approach revealed changes in the local epiproteome of a promoter during activation of transcription. CRISPR-ChAP-MS thus has broad applications for discovering molecular components and dynamic regulation of any in vivo activity at a given chromosomal location.

• 出版日期2014-9