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Epitope Tagging ChIP-Seq of DNA Binding Proteins Using CETCh-Seq
- 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.
- Subjects :
- 0303 health sciences
Cas9
genetic processes
Genomics
Computational biology
Biology
Genome
DNA sequencing
Epitope
03 medical and health sciences
0302 clinical medicine
Genome editing
CRISPR
natural sciences
Chromatin immunoprecipitation
030217 neurology & neurosurgery
030304 developmental biology
Subjects
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