Research output: Contribution to journal › Article › peer-review
Towards mapping the 3D genome through high speed single-molecule tracking of functional transcription factors in single living cells. / Wollman, Adam; Hedlund, Erik Goran; Shashkova, Sviatlana; Leake, Mark Christian.
In: Methods, 26.06.2019.Research output: Contribution to journal › Article › peer-review
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TY - JOUR
T1 - Towards mapping the 3D genome through high speed single-molecule tracking of functional transcription factors in single living cells
AU - Wollman, Adam
AU - Hedlund, Erik Goran
AU - Shashkova, Sviatlana
AU - Leake, Mark Christian
PY - 2019/6/26
Y1 - 2019/6/26
N2 - 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.
AB - 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.
KW - Gene regulation
KW - Single-molecule
KW - Super-resolution
KW - Transcription
KW - Transcription factors
KW - Yeast genome
UR - http://www.scopus.com/inward/record.url?scp=85068189701&partnerID=8YFLogxK
U2 - 10.1016/j.ymeth.2019.06.021
DO - 10.1016/j.ymeth.2019.06.021
M3 - Article
JO - Methods
JF - Methods
SN - 1046-2023
ER -