TY - CHAP
T1 - Chromatin immunoprecipitation in early mouse embryos
AU - García-González, Estela G.
AU - Roque-Ramirez, Bladimir
AU - Palma-Flores, Carlos
AU - Hernández-Hernández, J. Manuel
N1 - Publisher Copyright:
© 2018, Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2018
Y1 - 2018
N2 - Epigenetic regulation is achieved at many levels by different factors such as tissue-specific transcription factors, members of the basal transcriptional apparatus, chromatin-binding proteins, and noncoding RNAs. Importantly, chromatin structure dictates the availability of a specific genomic locus for transcriptional activation as well as the efficiency with which transcription can occur. Chromatin immunoprecipitation (ChIP) is a method that allows elucidating gene regulation at the molecular level by assessing if chromatin modifications or proteins are present at a specific locus. Initially, the majority of ChIP experiments were performed on cultured cell lines and more recently this technique has been adapted to a variety of tissues in different model organisms. Using ChIP on mouse embryos, it is possible to document the presence or absence of specific proteins and chromatin modifications at genomic loci in vivo during mammalian development and to get biological meaning from observations made on tissue culture analyses. We describe here a ChIP protocol on freshly isolated mouse embryonic somites for in vivo analysis of muscle specific transcription factor binding on chromatin. This protocol has been easily adapted to other mouse embryonic tissues and has also been successfully scaled up to perform ChIP-Seq.
AB - Epigenetic regulation is achieved at many levels by different factors such as tissue-specific transcription factors, members of the basal transcriptional apparatus, chromatin-binding proteins, and noncoding RNAs. Importantly, chromatin structure dictates the availability of a specific genomic locus for transcriptional activation as well as the efficiency with which transcription can occur. Chromatin immunoprecipitation (ChIP) is a method that allows elucidating gene regulation at the molecular level by assessing if chromatin modifications or proteins are present at a specific locus. Initially, the majority of ChIP experiments were performed on cultured cell lines and more recently this technique has been adapted to a variety of tissues in different model organisms. Using ChIP on mouse embryos, it is possible to document the presence or absence of specific proteins and chromatin modifications at genomic loci in vivo during mammalian development and to get biological meaning from observations made on tissue culture analyses. We describe here a ChIP protocol on freshly isolated mouse embryonic somites for in vivo analysis of muscle specific transcription factor binding on chromatin. This protocol has been easily adapted to other mouse embryonic tissues and has also been successfully scaled up to perform ChIP-Seq.
KW - Chromatin immunoprecipitation
KW - Embryo
KW - Epigenetics
KW - Gene regulation
KW - MyoD
KW - Myogenesis
KW - Myogenin
KW - Somites
UR - http://www.scopus.com/inward/record.url?scp=85044227799&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-7714-7_14
DO - 10.1007/978-1-4939-7714-7_14
M3 - Capítulo
C2 - 29564770
AN - SCOPUS:85044227799
T3 - Methods in Molecular Biology
SP - 145
EP - 155
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -