Epigenetic Regulation of Cardiac Differentiation of Embryonic Stem Cells and Tissues.
Specific gene transcription is a key biological process that underlies cell fate decision during embryonic development. The biological process is mediated by transcription factors which bind genomic regulatory regions including enhancers and promoters of cardiac constitutive genes. DNA is wrapped around histones that are subjected to chemical modifications. Modifications of histones further lead to repressed, activated or poised gene transcription, thus bringing another level of fine tuning regulation of gene transcription. Embryonic Stem cells (ES cells) recapitulate within embryoid bodies (i.e., cell aggregates) or in 2D culture the early steps of cardiac development. They provide in principle enough material for chromatin immunoprecipitation (ChIP), a technology broadly used to identify gene regulatory regions. Furthermore, human ES cells represent a human cell model of cardiogenesis. At later stages of development, mouse embryonic tissues allow for investigating specific epigenetic landscapes required for determination of cell identity. Herein, we describe protocols of ChIP, sequential ChIP followed by PCR or ChIP-sequencing using ES cells, embryoid bodies and cardiac specific embryonic regions. These protocols allow to investigating the epigenetic regulation of cardiac gene transcription.
INSERM UMR-910, Faculté de Médecine, Université Aix-Marseille.
Imen
I
Jebeniani
Institute of Physiological Chemistry, University Medical Center, Johannes Gutenberg University.
Julia
J
Leschik
INSERM UMR-910, Faculté de Médecine, Université Aix-Marseille; michel.puceat@inserm.fr.
Michel
M
Puceat
D000818:Animals; D002454:Cell Differentiation; D047369:Chromatin Immunoprecipitation; D044127:Epigenesis, Genetic; D006321:Heart; D006657:Histones; D000066449:Human Embryonic Stem Cells; D006801:Humans; D051379:Mice
D006657:Histones