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

STEM CELL TRANSCRIPTIONAL NETWORKS: METHODS AND PROTOCOLS, 2ND EDITION(2020)

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摘要
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.
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关键词
CRISPR, Epitope tagging, Gene regulation, Genomics, Transcription factors, ChIP-seq, CETCh-seq
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