mScarlet3: a brilliant and fast-maturing red fluorescent protein

被引：53

作者：

Gadella, Theodorus W. J. ^{[1
,2
]}

van Weeren, Laura ^{[1
,2
]}

Stouthamer, Jente ^{[1
,2
]}

Hink, Mark A. ^{[1
,2
]}

Wolters, Anouk H. G. ^{[3
]}

Giepmans, Ben N. G. ^{[3
]}

Aumonier, Sylvain ^{[4
]}

Dupuy, Jerome ^{[4
]}

Royant, Antoine ^{[4
,5
]}

机构：

[1] Univ Amsterdam, Swammerdam Inst Life Sci, Sect Mol Cytol, Amsterdam, Netherlands

[2] Univ Amsterdam, Swammerdam Inst Life Sci, Van Leeuwenhoek Ctr Adv Microscopy, Amsterdam, Netherlands

[3] Univ Groningen UMC Groningen, Biomed Sci Cells & Syst, Groningen, Netherlands

[4] Univ Grenoble Alpes, CNRS, CEA, Inst Biol Structurale, Grenoble, France

[5] European Synchrotron Radiat Facil, Grenoble, France

来源：

NATURE METHODS | 2023年 / 20卷 / 04期

关键词：

MONOMERIC RED; FEATURES; ORANGE;

D O I：

10.1038/s41592-023-01809-y

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We report the evolution of mScarlet3, a cysteine-free monomeric red fluorescent protein with fast and complete maturation, as well as record brightness, quantum yield (75%) and fluorescence lifetime (4.0 ns). The mScarlet3 crystal structure reveals a barrel rigidified at one of its heads by a large hydrophobic patch of internal residues. mScarlet3 behaves well as a fusion tag, displays no apparent cytotoxicity and it surpasses existing red fluorescent proteins as a Forster resonance energy transfer acceptor and as a reporter in transient expression systems. Two mScarlet variants with high brightness and fast maturation times have been evolved. These variants behave favorably as fusion tags and Forster resonance energy transfer acceptors.

引用

页码：541 / +

页数：27

共 30 条

[1]

Aslanturk O. S., 2018, GENOTOXICITY PREDICT, DOI [10.5772/intechopen.71923, DOI 10.5772/INTECHOPEN.71923]

[2] Improving brightness and photostability of green and red fluorescent proteins for live cell imaging and FRET reporting [J].

Bajar, Bryce T. ;

Wang, Emily S. ;

Lam, Amy J. ;

Kim, Bongjae B. ;

Jacobs, Conor L. ;

Howe, Elizabeth S. ;

Davidson, Michael W. ;

Lin, Michael Z. ;

Chu, Jun .

SCIENTIFIC REPORTS, 2016, 6

[3] Multiparameter screening method for developing optimized red-fluorescent proteins [J].

Bindels, Daphne S. ;

Postma, Marten ;

Haarbosch, Lindsay ;

van Weeren, Laura ;

Gadella, Theodorus W. J., Jr. .

NATURE PROTOCOLS, 2020, 15 (02) :450-478

[4] MScarlet: A bright monomeric red fluorescent protein for cellular imaging [J].

Bindels D.S. ;

Haarbosch L. ;

Van Weeren L. ;

Postma M. ;

Wiese K.E. ;

Mastop M. ;

Aumonier S. ;

Gotthard G. ;

Royant A. ;

Hink M.A. ;

Gadella T.W.J., Jr. .

Nature Methods, 2017, 14 (1) :53-56

[5] A monomeric red fluorescent protein [J].

Campbell, RE ;

Tour, O ;

Palmer, AE ;

Steinbach, PA ;

Baird, GS ;

Zacharias, DA ;

Tsien, RY .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (12) :7877-7882

[6] CrystalDirect: a new method for automated crystal harvesting based on laser-induced photoablation of thin films [J].

Cipriani, Florent ;

Roewer, Martin ;

Landret, Christophe ;

Zander, Ulrich ;

Felisaz, Franck ;

Marquez, Jose Antonio .

ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 2012, 68 :1393-1399

[7] A palette of fluorescent proteins optimized for diverse cellular environments [J].

Costantini, Lindsey M. ;

Baloban, Mikhail ;

Markwardt, Michele L. ;

Rizzo, Mark ;

Guo, Feng ;

Verkhusha, Vladislav V. ;

Snapp, Erik L. .

NATURE COMMUNICATIONS, 2015, 6

[8] Assessing the Tendency of Fluorescent Proteins to Oligomerize Under Physiologic Conditions [J].

Costantini, Lindsey M. ;

Fossati, Matteo ;

Francolini, Maura ;

Snapp, Erik Lee .

TRAFFIC, 2012, 13 (05) :643-649

[9] Reverse relation between cytotoxicity and Polyethylenimine/DNA ratio, the effect of using HEPES-buffered saline (HBS) medium in gene delivery [J].

Dabbaghi, Maryam ;

Hashemi, Khadijeh ;

Oskuee, Reza Kazemi ;

Afkhami-Goli, Amir .

TOXICOLOGY IN VITRO, 2022, 83

[10] ISPyB: an information management system for synchrotron macromolecular crystallography [J].

Delageniere, Solange ;

Brenchereau, Patrice ;

Launer, Ludovic ;

Ashton, Alun W. ;

Leal, Ricardo ;

Veyrier, Stephanie ;

Gabadinho, Jose ;

Gordon, Elspeth J. ;

Jones, Samuel D. ;

Levik, Karl Erik ;

McSweeney, Sean M. ;

Monaco, Stephanie ;

Nanao, Max ;

Spruce, Darren ;

Svensson, Olof ;

Walsh, Martin A. ;

Leonard, Gordon A. .

BIOINFORMATICS, 2011, 27 (22) :3186-3192

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