Translocation Biosensors - Cellular System Integrators to Dissect CRM1-Dependent Nuclear Export by Chemicogenomics.

Sensors
Verena FetzR H Stauber

Abstract

Fluorescent protein biosensors are powerful cellular systems biology tools for dissecting the complexity of cellular processes with high spatial and temporal resolution. As regulated nucleo-cytoplasmic transport is crucial for the modulation of numerous (patho)physiological cellular responses, a detailed understanding of its molecular mechanism would open up novel options for a rational manipulation of the cell. In contrast to genetic approaches, we here established and employed high-content cellular translocation biosensors applicable for dissecting nuclear export by chemicogenomics. A431 cell lines, stably expressing a translocation biosensor composed of glutathione S-transferase, GFP and a rational combination of nuclear import and export signals, were engineered by antibiotic selection and flow cytometry sorting. Using an optimized nuclear translocation algorithm, the translocation response could be robustly quantified on the Cellomics Arrayscan(®) VTI platform. Subsequent to assay optimization, the assay was developed into a higher density 384-well format high-content assay and employed for the screening of the 17K ChemBioNet compound collection. This library was selected on the basis of a genetic algorithm used to identif...Continue Reading

References

Apr 4, 1998·BioTechniques·R H StauberG N Pavlakis
Jan 3, 2001·Journal of Cellular Biochemistry·R H StauberO Rosorius
Jun 7, 2003·Developmental Cell·Mythili Suntharalingam, Susan R Wente
Jan 21, 2004·Nature Reviews. Cancer·Tweeny R KauPamela A Silver
May 8, 2004·Nature Reviews. Genetics·Markus Bredel, Edgar Jacoby
Sep 29, 2004·Trends in Cell Biology·Nima Mosammaparast, Lucy F Pemberton
Oct 12, 2004·FEBS Letters·Torsten MeissnerUwe Vinkemeier
Dec 17, 2004·Nature·Christopher Lipinski, Andrew Hopkins
Feb 11, 2005·Traffic·Lucy F Pemberton, Bryce M Paschal
May 24, 2005·Nature Cell Biology·Alexei ArnaoutovMary Dasso
Apr 28, 2006·Drug Discovery Today·Peter B Simpson, Keith A Wafford
Jun 3, 2006·Methods in Molecular Biology·Bohumil MacoUeli Aebi
Nov 18, 2006·Methods in Enzymology·Kenneth A GiulianoD Lansing Taylor
Feb 24, 2007·Current Opinion in Genetics & Development·Christopher R Brown, Pamela A Silver
Feb 24, 2007·Trends in Cell Biology·Saskia Hutten, Ralph H Kehlenbach
Jul 10, 2007·Cancer Research·Roland H StauberShirley K Knauer
Oct 6, 2007·Science·Roderick Y H LimUeli Aebi
Dec 12, 2007·Nature Reviews. Genetics·Suzanne Komili, Pamela A Silver
Mar 28, 2008·International Journal of Cancer. Journal International Du Cancer·Tim BeissertMartin Ruthardt
Sep 13, 2008·Trends in Cell Biology·Maximiliano A D'Angelo, Martin W Hetzer
Sep 18, 2008·Combinatorial Chemistry & High Throughput Screening·Michael WolffRalf Heilker
Nov 28, 2008·Assay and Drug Development Technologies·Sam MichaelAnton Simeonov
Jan 9, 2009·International Journal of Cancer. Journal International Du Cancer·Verena FetzShirley K Knauer
Jan 28, 2009·Genes & Development·Oliver H KrämerThorsten Heinzel

❮ Previous
Next ❯

Citations

Mar 3, 2011·Cell Death & Disease·S K KnauerR H Stauber
Jul 16, 2010·Molecular and Cellular Neurosciences·Negusse HabtemichaelWolf J Mann
Mar 23, 2011·Traffic·Carolin BierRoland H Stauber
Jun 18, 2016·Cellular and Molecular Life Sciences : CMLS·Iraia García-SantistebanJose A Rodríguez
May 16, 2013·Sensors·Nicola S FracchiollaAgostino Cortelezzi
Oct 11, 2017·Journal of Molecular Cell Biology·Christian SchrenkShirley K Knauer
Nov 7, 2019·The Journal of Cell Biology·Panagiotis MistriotisKonstantinos Konstantopoulos
Jan 31, 2015·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Désirée WünschRoland H Stauber
Feb 27, 2015·Biological Chemistry·Désirée WünschShirley K Knauer
Nov 19, 2010·The Journal of Biological Chemistry·Carolin BierRoland H Stauber
Sep 24, 2014·Beilstein Journal of Nanotechnology·Dominic DocterRoland H Stauber
Jan 10, 2018·Sensors·Mohammad RizwanMinhaz Uddin Ahmed
Mar 9, 2021·Acta Crystallographica. Section F, Structural Biology Communications·Alaa ShaikhqasemRalf Ficner
Jun 26, 2020·Journal of Medicinal Chemistry·Alaa ShaikhqasemRalf Ficner

❮ Previous
Next ❯

Methods Mentioned

BETA
biosensor
biosensors
fluorescence microscopy
Nucleotide Exchange
transfection
RevNES-biosensor
fluorescence activated cell sorting
Assay
ELISA

Software Mentioned

ArrayScan
ArrayScan ® view
Cellomics
AxioVision
ArrayScan view
Cellomics Arrayscan

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.