DamID to Map Genome-Protein Interactions in Preimplantation Mouse Embryos.
Investigating the chromatin landscape of the early mammalian embryo is essential to understand how epigenetic mechanisms may direct reprogramming and cell fate allocation. Genome-wide analyses of the epigenome in preimplantation mouse embryos have recently become available, thanks to the development of low-input protocols. DNA adenine methyltransferase identification (DamID) enables the investigation of genome-wide protein-DNA interactions without the requirement of specific antibodies. Most importantly, DamID can be robustly applied to single cells. Here we describe the protocol for performing DamID in single oocytes and mouse preimplantation embryos, as well as single blastomeres, using a Dam-LaminB1 fusion to generate high-resolution lamina-associated domain (LAD) maps. This low-input method can be adapted for other proteins of interest to faithfully profile their genomic interaction, allowing us to interrogate the chromatin dynamics and nuclear organization during the early mammalian development.
['Pal|Mrinmoy|M|', 'Kind|Jop|J|', 'Torres-Padilla|Maria-Elena|ME|']
D000818:Animals D001755:Blastocyst D002478:Cells, Cultured D046149:Embryo Culture Techniques D016678:Genome D023281:Genomics D059014:High-Throughput Nucleotide Sequencing D051379:Mice D008810:Mice, Inbred C57BL D009865:Oocytes D059010:Single-Cell Analysis D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific)
Mouse embryo LADs Nuclear organization Low-input DamID Single-cell genomics