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

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JournalMethods
DateAccepted/In press - 22 Jun 2019
DateE-pub ahead of print - 26 Jun 2019
DatePublished (current) - 26 Jun 2019
Number of pages8
Early online date26/06/19
Original languageEnglish

Abstract

How 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.

    Research areas

  • Gene regulation, Single-molecule, Super-resolution, Transcription, Transcription factors, Yeast genome

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