title: A CRISPR-based approach for proteomic analysis of a single genomic locus.
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 (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.
authors.affiliation: University of Arkansas for Medical Sciences; Department of Biochemistry and Molecular Biology; Little Rock, AR USA.
authors.forename: Zachary J
authors.initials: ZJ
authors.lastname: Waldrip
authors.affiliation: University of Arkansas for Medical Sciences; Department of Biochemistry and Molecular Biology; Little Rock, AR USA.
authors.forename: Stephanie D
authors.initials: SD
authors.lastname: Byrum
authors.affiliation: University of Arkansas for Medical Sciences; Department of Biochemistry and Molecular Biology; Little Rock, AR USA.
authors.forename: Aaron J
authors.initials: AJ
authors.lastname: Storey
authors.affiliation: University of Arkansas for Medical Sciences; Department of Biochemistry and Molecular Biology; Little Rock, AR USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Gao
authors.affiliation: University of Arkansas for Medical Sciences; Department of Biochemistry and Molecular Biology; Little Rock, AR USA.
authors.forename: Alicia K
authors.initials: AK
authors.lastname: Byrd
authors.affiliation: University of Arkansas for Medical Sciences; Department of Biochemistry and Molecular Biology; Little Rock, AR USA.
authors.forename: Samuel G
authors.initials: SG
authors.lastname: Mackintosh
authors.affiliation: University of Arkansas for Medical Sciences; Department of Biochemistry and Molecular Biology; Little Rock, AR USA.
authors.forename: Wayne P
authors.initials: WP
authors.lastname: Wahls
authors.affiliation: Department of Pharmacology and Molecular Sciences; Johns Hopkins University School of Medicine; Baltimore, MD USA.
authors.forename: Sean D
authors.initials: SD
authors.lastname: Taverna
authors.affiliation: University of Arkansas for Medical Sciences; Department of Biochemistry and Molecular Biology; Little Rock, AR USA.
authors.forename: Kevin D
authors.initials: KD
authors.lastname: Raney
authors.affiliation: University of Arkansas for Medical Sciences; Department of Biochemistry and Molecular Biology; Little Rock, AR USA.
authors.forename: Alan J
authors.initials: AJ
authors.lastname: Tackett
mesh_terms: D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D056426:Genetic Loci; D006657:Histones / Q000378:metabolism*; D013058:Mass Spectrometry; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000235:genetics / Q000378:metabolism*; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D002843:Chromatin; D006657:Histones; D020543:Proteome; D000094704:RNA, Guide, CRISPR-Cas Systems; D029701:Saccharomyces cerevisiae Proteins
keywords: affinity purification; epigenetics; epiproteome; histone; posttranslational modification; proteomics