DamID to Map Genome-Protein Interactions in Preimplantation Mouse Embryos

Mrinmoy Pal; Jop Kind; Maria-Elena Torres-Padilla

doi:10.1007/978-1-0716-0958-3_18

DamID to Map Genome-Protein Interactions in Preimplantation Mouse Embryos

Mrinmoy Pal, Jop Kind, Maria-Elena Torres-Padilla

Hubrecht Institute

Research output: Contribution to journal/periodical › Article › Scientific › peer-review

3 Citations (Scopus)

Abstract

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.

Original language	English
Pages (from-to)	265-282
Number of pages	18
Journal	Methods in molecular biology (Clifton, N.J.)
Volume	2214
DOIs	https://doi.org/10.1007/978-1-0716-0958-3_18
Publication status	Published - 2020

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title = "DamID to Map Genome-Protein Interactions in Preimplantation Mouse Embryos",

abstract = "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.",

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AU - Pal, Mrinmoy

AU - Kind, Jop

AU - Torres-Padilla, Maria-Elena

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AB - 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.

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JO - Methods in molecular biology (Clifton, N.J.)

JF - Methods in molecular biology (Clifton, N.J.)

ER -

DamID to Map Genome-Protein Interactions in Preimplantation Mouse Embryos

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