mGreenLantern: a bright monomeric fluorescent protein with rapid expression and cell filling properties for neuronal imaging.

Proceedings of the National Academy of Sciences of the United States of America
Benjamin C CampbellGregory A Petsko

Abstract

Although ubiquitous in biological studies, the enhanced green and yellow fluorescent proteins (EGFP and EYFP) were not specifically optimized for neuroscience, and their underwhelming brightness and slow expression in brain tissue limits the fidelity of dendritic spine analysis and other indispensable techniques for studying neurodevelopment and plasticity. We hypothesized that EGFP's low solubility in mammalian systems must limit the total fluorescence output of whole cells, and that improving folding efficiency could therefore translate into greater brightness of expressing neurons. By introducing rationally selected combinations of folding-enhancing mutations into GFP templates and screening for brightness and expression rate in human cells, we developed mGreenLantern, a fluorescent protein having up to sixfold greater brightness in cells than EGFP. mGreenLantern illuminates neurons in the mouse brain within 72 h, dramatically reducing lag time between viral transduction and imaging, while its high brightness improves detection of neuronal morphology using widefield, confocal, and two-photon microscopy. When virally expressed to projection neurons in vivo, mGreenLantern fluorescence developed four times faster than EYFP and ...Continue Reading

References

Dec 20, 1994·Proceedings of the National Academy of Sciences of the United States of America·R HeimR Y Tsien
Mar 6, 1997·Biochemical and Biophysical Research Communications·Y KimataK Kohno
May 29, 2003·BMC Biotechnology·Otilia Zapata-Hommer, Oliver Griesbeck
Nov 22, 2005·Nature Methods·Nathan C ShanerRoger Y Tsien
Dec 22, 2005·Nature Biotechnology·Jean-Denis PédelacqGeoffrey S Waldo
May 6, 2008·Nature Methods·Nathan C ShanerRoger Y Tsien
Apr 16, 2009·The Journal of Physical Chemistry. B·Valentin LulevichGang-Yu Liu
Apr 21, 2009·Methods in Molecular Biology·Alan R HarveyChristiaan N Levelt
Sep 23, 2009·Chemical Society Reviews·Timothy D Craggs
Apr 5, 2011·Analytical Biochemistry·Ryo IizukaTakashi Funatsu
Mar 22, 2012·Nature Communications·Joachim GoedhartAntoine Royant
Sep 11, 2012·Nature Methods·Amy J LamMichael Z Lin
Oct 13, 2012·Nature Protocols·Ali ErtürkHans-Ulrich Dodt
Jan 29, 2013·International Review of Cell and Molecular Biology·Olesya V StepanenkoKonstantin K Turoverov
Jul 23, 2013·Nature·Tsai-Wen ChenDouglas S Kim
Jan 7, 2014·Science China. Life Sciences·JingJin DingFeng Shao
Jun 18, 2014·Nature Chemical Biology·Kevin M Dean, Amy E Palmer
Oct 28, 2014·PloS One·Olesya V StepanenkoKonstantin K Turoverov
May 30, 2015·ENeuro·Cynthia SoderblomPantelis Tsoulfas
May 31, 2016·Nature Methods·Paula J CranfillDavid W Piston
Nov 5, 2016·Molecular Biology of the Cell·Jennifer K HeppertBob Goldstein
Nov 7, 2016·Trends in Biochemical Sciences·Erik A RodriguezRoger Y Tsien
Nov 22, 2016·Nature Methods·Daphne S BindelsTheodorus W J Gadella
Oct 7, 2017·Cell·Douglas S Richardson, Jeff W Lichtman
Jul 10, 2018·Biomedical Optics Express·Antony OrthBrant C Gibson
Jul 13, 2018·Frontiers in Neuroscience·Peter VerstraelenWinnok H De Vos
Aug 3, 2018·Current Protocols in Cell Biology·Shoh M AsanoEdward S Boyden
Oct 21, 2018·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Zimei WangMurray G Blackmore

❮ Previous
Next ❯

Citations

Jan 3, 2021·Trends in Cell Biology·Julien CassereauU Valentin Nägerl
May 1, 2021·Pharmaceuticals·Jung Hak KimHong Jun Lee
Jun 15, 2021·Genesis : the Journal of Genetics and Development·Kalene R JassoJeremy C McIntyre
Jul 6, 2021·STAR Protocols·Antoine Gaudreau-LapierreLaura Trinkle-Mulcahy
Jul 3, 2021·International Journal of Molecular Sciences·Paweł MatrybaJakub Gołąb
Sep 15, 2021·Experimental Neurology·Murray BlackmorePantelis Tsoulfas
Jun 26, 2021··Jean-Claude BéïqueAntoine Gaudreau-Lapierre

❮ Previous
Next ❯

Methods Mentioned

BETA
transgenic
transfection
confocal microscopy
PCR
light sheet microscopy
gel filtration
fluorescence microscopy
biosensors
Fluorescence
Protein Assay

Software Mentioned

Fluoview
Fiji
NIS
tdTomato
mScarlet
Bio
Clover
Elements AR
Rad CFX96
ImageJ

Related Concepts

Related Feeds

Advanced Imaging of Cellular Signaling

Cell signaling is a vital mechanism for communication within cells and outside with the environment. Several different signaling pathways have been found and advanced imaging techniques are being developed to visualize the molecules involved in these signaling pathways. Find the latest research in advanced imaging of cellular signaling here.