Presentation Title

Characterizing Nucleolar Associated Heterochromatin in Mouse Cells Using 3D-FISH

Faculty Mentor

Paul D. Kaufman

Start Date

18-11-2017 10:00 AM

End Date

18-11-2017 11:00 AM

Location

BSC-Ursa Minor 35

Session

Poster 1

Type of Presentation

Poster

Subject Area

biological_agricultural_sciences

Abstract

Kevin Ou, Anastassia Vertii, Paul D. Kaufman

Department of Molecular, Cell and Cancer Biology

Characterizing Nucleolar Associated Heterochromatin in Mouse Cells Using 3D-FISH

A complete understanding of epigenetics will require knowledge about the relationships between nuclear organization and gene regulation. Recent research has shown that localization is critical for gene regulation of heterochromatin, the more condensed and transcriptionally repressed chromatin that frequently tethers to either the nuclear lamina or nucleolus. Additionally, although it is known that perinucleolar heterochromatin is particularly dynamic in developing mouse embryos, it has not been mapped systematically. Thus, we seek to map heterochromatin changes in mouse embryonic stem cells (mESC) before and during differentiation into neural progenitor cells. As a first step towards this, we developed methods for isolating, deep sequencing, and analysis of associated DNA of the nucleoli in mouse embryonic fibroblasts (MEFs). These studies revealed a new class of nucleolar associated domain (NAD) of heterochromatin that preferentially localizes to the nucleolus but not to the nuclear lamina: class II NADs. We confirmed the sequencing results by performing 3D fluorescence in situ hybridization (FISH) with class II NAD probes and quantifying their association with the nucleolus. Our results altogether provide convincing evidence of the existence of class II NADs in mouse cells and will be used to characterize NAD changes during differentiation.

This project was supported by NIH Grant U01 DA040588 as part of the 4DN Network.

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Nov 18th, 10:00 AM Nov 18th, 11:00 AM

Characterizing Nucleolar Associated Heterochromatin in Mouse Cells Using 3D-FISH

BSC-Ursa Minor 35

Kevin Ou, Anastassia Vertii, Paul D. Kaufman

Department of Molecular, Cell and Cancer Biology

Characterizing Nucleolar Associated Heterochromatin in Mouse Cells Using 3D-FISH

A complete understanding of epigenetics will require knowledge about the relationships between nuclear organization and gene regulation. Recent research has shown that localization is critical for gene regulation of heterochromatin, the more condensed and transcriptionally repressed chromatin that frequently tethers to either the nuclear lamina or nucleolus. Additionally, although it is known that perinucleolar heterochromatin is particularly dynamic in developing mouse embryos, it has not been mapped systematically. Thus, we seek to map heterochromatin changes in mouse embryonic stem cells (mESC) before and during differentiation into neural progenitor cells. As a first step towards this, we developed methods for isolating, deep sequencing, and analysis of associated DNA of the nucleoli in mouse embryonic fibroblasts (MEFs). These studies revealed a new class of nucleolar associated domain (NAD) of heterochromatin that preferentially localizes to the nucleolus but not to the nuclear lamina: class II NADs. We confirmed the sequencing results by performing 3D fluorescence in situ hybridization (FISH) with class II NAD probes and quantifying their association with the nucleolus. Our results altogether provide convincing evidence of the existence of class II NADs in mouse cells and will be used to characterize NAD changes during differentiation.

This project was supported by NIH Grant U01 DA040588 as part of the 4DN Network.