RT Journal Article
SR Electronic
T1 Cell Type Specific Survey of Epigenetic Modifications by Tandem Chromatin Immunoprecipitation Sequencing
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 163568
DO 10.1101/163568
A1 Mari Mito
A1 Mitsutaka Kadota
A1 Kaori Tanaka
A1 Yasuhide Furuta
A1 Kuniya Abe
A1 Shintaro Iwasaki
A1 Shinichi Nakagawa
YR 2017
UL http://biorxiv.org/content/early/2017/07/14/163568.abstract
AB Background The nervous system of higher eukaryotes is composed of numerous types of neurons and glia that together orchestrate complex neuronal responses. However, this complex pool of cells typically poses analytical challenges in investigating gene expression profiles and their epigenetic basis for specific cell types. Here, we developed a novel method that enables cell type-specific analyses of epigenetic modifications using tandem chromatin immunoprecipitation sequencing (tChIP-Seq).Results FLAG-tagged histone H2B, a constitutive chromatin component, was first expressed in Camk2a-positive pyramidal cortical neurons and used to purify chromatin in a cell type-specific manner. Subsequent chromatin immunoprecipitation using antibodies against H3K4me3—an active promoter mark—allowed us to survey neuron-specific coding and non-coding transcripts. Indeed, tChIP-Seq identified hundreds of genes associated with neuronal functions and genes with unknown functions expressed in cortical neurons.Conclusions tChIP-Seq thus provides a versatile approach to investigating the epigenetic modifications of particular cell types in vivo.