Imaging of native transcription factors and histone phosphorylation at high resolution in live cells

Conic, Sascha and Desplancq, Dominique and Ferrand, Alexia and Fischer, Veronique and Heyer, Vincent and Reina San Martin, Bernardo and Pontabry, Julien and Oulad-Abdelghani, Mustapha and Babu N, Kishore and Wright, Graham D. and Molina, Nacho and Weiss, Etienne and Tora, László. (2018) Imaging of native transcription factors and histone phosphorylation at high resolution in live cells. The Journal of cell biology, 217 (4). pp. 1537-1552.

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Official URL: https://edoc.unibas.ch/64879/

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Fluorescent labeling of endogenous proteins for live-cell imaging without exogenous expression of tagged proteins or genetic manipulations has not been routinely possible. We describe a simple versatile antibody-based imaging approach (VANIMA) for the precise localization and tracking of endogenous nuclear factors. Our protocol can be implemented in every laboratory allowing the efficient and nonharmful delivery of organic dye-conjugated antibodies, or antibody fragments, into different metazoan cell types. Live-cell imaging permits following the labeled probes bound to their endogenous targets. By using conventional and super-resolution imaging we show dynamic changes in the distribution of several nuclear transcription factors (i.e., RNA polymerase II or TAF10), and specific phosphorylated histones (γH2AX), upon distinct biological stimuli at the nanometer scale. Hence, considering the large panel of available antibodies and the simplicity of their implementation, VANIMA can be used to uncover novel biological information based on the dynamic behavior of transcription factors or posttranslational modifications in the nucleus of single live cells.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Services Biozentrum > Imaging Core Facility (Biehlmaier)
UniBasel Contributors:Loynton-Ferrand, Alexia
Item Type:Article, refereed
Article Subtype:Research Article
Note:Publication type according to Uni Basel Research Database: Journal article
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Last Modified:27 Apr 2020 10:03
Deposited On:27 Apr 2020 10:03

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