Epitope Tagging ChIP-Seq of DNA Binding Proteins Using CETCh-Seq

Methods Mol Biol. 2020:2117:3-34. doi: 10.1007/978-1-0716-0301-7_1.

Abstract

Chromatin immunoprecipitation followed by next-generation DNA sequencing (ChIP-seq) has been used to identify transcription factor (TF) binding proteins throughout the genome. Unfortunately, this approach traditionally requires commercially available, ChIP-seq grade antibodies that frequently fail to generate acceptable datasets. To obtain data for the many TFs for which there is no appropriate antibody, we recently developed a new method for performing ChIP-seq by epitope tagging endogenous TFs using CRISPR/Cas9 genome editing technology (CETCh-seq). Here, we describe our general protocol of CETCh-seq for both adherent and nonadherent cell lines using a commercially available FLAG antibody.

Keywords: CETCh-seq; CRISPR; ChIP-seq; Epitope tagging; Gene regulation; Genomics; Transcription factors.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Binding Sites
  • CRISPR-Cas Systems
  • Cell Adhesion
  • Chromatin Immunoprecipitation Sequencing
  • Epitopes / metabolism*
  • Gene Editing
  • Hep G2 Cells
  • Humans
  • Protein Binding
  • Transcription Factors / analysis*
  • Transcription Factors / genetics*

Substances

  • Epitopes
  • Transcription Factors