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

Authors :
Laurel A. Brandsmeier
Mark Mackiewicz
Amy S. Nesmith
Eric M. Mendenhall
Richard M. Myers
Sarah Meadows
Michael J Betti
Kimberly M. Newberry
E. Christopher Partridge
Source :
Methods in Molecular Biology ISBN: 9781071603000
Publication Year :
2020
Publisher :
Springer US, 2020.

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.

Details

ISBN :
978-1-07-160300-0
ISBNs :
9781071603000
Database :
OpenAIRE
Journal :
Methods in Molecular Biology ISBN: 9781071603000
Accession number :
edsair.doi...........384c984eb0a3c9a205c14ac264c72f70
Full Text :
https://doi.org/10.1007/978-1-0716-0301-7_1