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VisuLive SIGNED

Quantitative Nanoscale Visualization of Macromolecular Complexes in Live Cells using Genetic Code Expansion and High-Resolution Imaging

Total Cost €

EC-Contrib. €

Partnership

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Outcomes and
results

VisuLive project word cloud

Explore the words cloud of the VisuLive project. It provides you a very rough idea of what is the project "VisuLive" about.

labeling visualizing dynamin spatiotemporal complexes fluorescence implications mechanistic techniques escrt demonstrated speed dyes innovative first shortcoming tags fluorescent function bulky substituting expansion high organization cell membrane decipher markedly code kinetics rely live principles structural quantitative mechanical formulate cellular gfp possess super bioorthogonal contribution superior photophysical rate protein fission biology proteins context distinguished physiological cells time nanoscale invasive scope stresses smaller label obtain close genetic milliseconds attach broad resolution microscopy imaging

Project "VisuLive" data sheet

The following table provides information about the project.

Coordinator	BEN-GURION UNIVERSITY OF THE NEGEV Organization address address: . city: BEER SHEVA postcode: 84105 website: www.bgu.ac.il contact info title: n.a. name: n.a. surname: n.a. function: n.a. email: n.a. telephone: n.a. fax: n.a.
Coordinator Country	Israel [IL]
Total cost	1˙625˙000 €
EC max contribution	1˙625˙000 € (100%)
Programme	1. H2020-EU.1.1. (EXCELLENT SCIENCE - European Research Council (ERC))
Code Call	ERC-2014-STG
Funding Scheme	ERC-STG
Starting year	2015
Duration (year-month-day)	from 2015-04-01 to 2020-03-31

Partnership

Take a look of project's partnership.

#	participants	country	role	EC contrib. [€]
1	BEN-GURION UNIVERSITY OF THE NEGEV BEN-GURION UNIVERSITY OF THE NEGEV Organization address address: . city: BEER SHEVA postcode: 84105 website: www.bgu.ac.il contact info title: n.a. name: n.a. surname: n.a. function: n.a. email: n.a. telephone: n.a. fax: n.a.	IL (BEER SHEVA)	coordinator	1˙625˙000.00

Map

Project objective

High-resolution fluorescence imaging, including super-resolution microscopy and high-speed live cell imaging, are used to obtain quantitative information on the structural organization and kinetics of cellular processes. The contribution of these high-resolution techniques to cell biology was recently demonstrated for dynamin- and ESCRT-driven membrane fission in cells. While they advance our knowledge on membrane fission these techniques do not provide the quantitative information needed to formulate a mechanical understanding of membrane fission in a physiological context, a shortcoming that stresses the need to increase the spatiotemporal resolution and improve the live cell capabilities of these techniques. Substituting the bulky fluorescent protein tags (such as GFP) currently used in live-cell applications with much smaller fluorescent dyes that possess superior photophysical characteristics will markedly improve these advanced imaging techniques. Genetic code expansion and bioorthogonal labeling offer, for the first time, a non-invasive way to specifically attach such fluorescent dyes to proteins in live cells. I, therefore, propose to develop an innovative approach to label cellular proteins with fluorescent dyes via genetic code expansion for quantitative high-resolution live cell imaging of cellular protein complexes. By applying this approach to three distinguished high-resolution methodologies and by visualizing membrane fission in distinct cellular processes in live cells at milliseconds rate and at nanoscale resolution, we aim to decipher the mechanistic principles of membrane fission in cells. As numerous cellular processes rely on membrane fission for their function, such an understanding will have a broad impact on cell biology. The implications of this study reach beyond the scope of membrane fission by offering a new approach to study cellular processes at close-to-real conditions in live cells and at nanoscale resolution.

Publications

List of publications.
year	authors and title	journal	last update
2016	Sarit Cohen, Eyal Arbely Single-Plasmid-Based System for Efficient Noncanonical Amino Acid Mutagenesis in Cultured Mammalian Cells published pages: 1008-1011, ISSN: 1439-4227, DOI: 10.1002/cbic.201500681	ChemBioChem 17/11	2019-05-29
2017	Tomer Schvartz, Noa Aloush, Inna Goliand, Inbar Segal, Dikla Nachmias, Eyal Arbely, Natalie Elia Direct fluorescent-dye labeling of Î±-tubulin in mammalian cells for live cell and superresolution imaging published pages: 2747-2756, ISSN: 1059-1524, DOI: 10.1091/mbc.E17-03-0161	Molecular Biology of the Cell 28/21	2019-05-29

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The information about "VISULIVE" are provided by the European Opendata Portal: CORDIS opendata.