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Towards mapping the 3D genome through high speed single-molecule tracking of functional transcription factors in single living cells

Lookup NU author(s): Dr Adam WollmanORCiD

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).

Abstract

© 2019 The AuthorsHow genomic DNA is organized in the nucleus is a long-standing question. We describe a single-molecule bioimaging method utilizing super-localization precision coupled to fully quantitative image analysis tools, towards determining snapshots of parts of the 3D genome architecture of model eukaryote budding yeast Saccharomyces cerevisiae with exceptional millisecond time resolution. We employ astigmatism imaging to enable robust extraction of 3D position data on genomically encoded fluorescent protein reporters that bind to DNA. Our relatively straightforward method enables snippets of 3D architectures of likely single genome conformations to be resolved captured via DNA-sequence specific binding proteins in single functional living cells.

Publication metadata

Author(s): Wollman AJM, Hedlund EG, Shashkova S, Leake MC

Publication type: Article

Publication status: Published

Journal: Methods

Year: 2020

Volume: 170

Pages: 82-89

Print publication date: 01/01/2020

Online publication date: 26/06/2019

Acceptance date: 22/06/2019

Date deposited: 10/02/2020

ISSN (print): 1046-2023

ISSN (electronic): 1095-9130

Publisher: Academic Press Inc.

URL: https://doi.org/10.1016/j.ymeth.2019.06.021

DOI: 10.1016/j.ymeth.2019.06.021

PubMed id: 31252059

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