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mNeonGreen

a.k.a. mNG

mNeonGreen is a basic (constitutively fluorescent) green/yellow fluorescent protein published in 2013, derived from Branchiostoma lanceolatum. It is reported to be a very rapidly-maturing monomer with moderate acid sensitivity.
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Oligomerization Organism Molecular Weight Cofactor
Monomer Branchiostoma lanceolatum 26.6 kDa -

FPbase ID: ZRKRV

Attributes

Ex λ Em λ EC (M-1 cm-1) QY Brightness pKa Maturation (min) Lifetime (ns)
506 517 116,000 0.8 92.8 5.7 10.0 3.1
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mNeonGreen OSER Measurements

% Normal Cells OSER/NE ratio Cell Type Reference
90.4 ± 2.1 (10000 cells) - HeLa Shaner et al. (2013)
90.4 ± 2.1 (10000 cells) - HeLa Cranfill et al. (2016)

Photostability

t1/2 (s) Power Light Mode In Cell Fusion ˚C Reference
158.0 4.3 (mW) Arc-lamp Widefield H2B Shaner et al. (2013)
150.0 7.4 (W/cm2) Laser Spinning Disc Confocal HeLa (fixed) TPM2 25.0 Ivorra-Molla et al. (2023)
A caution on interpretation of photostability measurements
Add photostability info

mNeonGreen Sequence

BLAST

mNeonGreen was derived from dLanYFP with the following mutations: F15I/R25Q/Q56H/F67Y/K79V/S100V/F115A/V140R/T141S/M143K/L144T/D156K/T158S/Q168R/I185Y/F192Y
amino acid numbers relative to LanYFP. show relative to dLanYFP

MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDGASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIISTFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQKAFTDVMGMDELYK
GenBank: AGG56535
IPG: 34051755

Structure

Deposited: ,
Chromophore:

Excerpts

mNeonGreen is the brightest monomeric green or yellow fluorescent protein yet described to our knowledge, performs exceptionally well as a fusion tag for traditional imaging as well as stochastic single-molecule superresolution imaging and is an excellent fluorescence resonance energy transfer (FRET) acceptor for the newest cyan fluorescent proteins.

Shaner et al. (2013)

... as it has so little sequence identity in common with other frequently used fluorescent proteins, mNeonGreen will be an attractive target for antibody development and should be amenable to orthogonal immunoprecipitation experiments along with jellyfish-derived and coral-derived fluorescent proteins.

Shaner et al. (2013)

Notably, when both mTurquoise2 and mNeonGreen donors were included simultaneously, double-labeled cells could be found, which appeared to be capable of dividing (Figure 4—figure supplement 1), indicating that FP labeling does not affect cell viability in diploid CRISPIEd cells

Zhong et al. (2021)

Primary Reference

A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum

Shaner Nc, Lambert Gg, Chammas A, Ni Y, Cranfill Pj, Baird Ma, Sell Br, Allen Jr, Day Rn, Israelsson M, Davidson Mw, Wang J

(2013). Nature Methods, 10(5) , 407-409. doi: 10.1038/nmeth.2413. Article   Pubmed

Additional References

  1. High-fidelity, efficient, and reversible labeling of endogenous proteins using CRISPR-based designer exon insertion

    Zhong H, Ceballos Cc, Massengill Ci, Muniak Ma, Ma L, Qin M, Petrie Sk, Mao T

    (2021). eLife, 10, . doi: 10.7554/elife.64911. Article

  2. Comparing the performance of mScarlet-I, mRuby3, and mCherry as FRET acceptors for mNeonGreen

    Mccullock Tw, Maclean Dm, Kammermeier Pj

    (2020). PLOS ONE, 15(2) , e0219886. doi: 10.1371/journal.pone.0219886. Article   Pubmed

  3. Identification and Spectroscopic Characterization of mNeonGreen as a pH-Dependent, Turn-on Fluorescent Protein Sensor for Chloride

    Tutol J, Kam H, Dodani S

    (2019). ChemBioChem, , . doi: 10.1002/cbic.201900147. Article   Pubmed

  4. I see the light! Fluorescent proteins suitable for cell wall/apoplast targeting in Nicotiana benthamiana leaves

    Stoddard A, Rolland V

    (2019). Plant Direct, 3(1) , e00112. doi: 10.1002/pld3.112. Article   Pubmed

  5. Optimization of sample preparation and green color imaging using the mNeonGreen fluorescent protein in bacterial cells for photoactivated localization microscopy

    Stockmar I, Feddersen H, Cramer K, Gruber S, Jung K, Bramkamp M, Shin Jy

    (2018). Scientific Reports, 8(1) , 10137. doi: 10.1038/s41598-018-28472-0. Article   Pubmed

  6. Photophysical Behavior of mNeonGreen, an Evolutionarily Distant Green Fluorescent Protein

    Steiert F, Petrov Ep, Schultz P, Schwille P, Weidemann T

    (2018). Biophysical Journal, 114(10) , 2419-2431. doi: 10.1016/j.bpj.2018.04.013. Article   Pubmed

  7. Optimization of mNeonGreen for Homo sapiens increases its fluorescent intensity in mammalian cells

    Tanida-Miyake E, Koike M, Uchiyama Y, Tanida I

    (2018). PLOS ONE, 13(1) , e0191108. doi: 10.1371/journal.pone.0191108. Article   Pubmed

  8. Robust and Bright Genetically Encoded Fluorescent Markers for Highlighting Structures and Compartments in Mammalian Cells

    Chertkova Ao, Mastop M, Postma M, Van Bommel N, Van Der Niet S, Batenburg Kl, Joosen L, Gadella Twj, Okada Y, Goedhart J

    (2017). , , . doi: 10.1101/160374. Article

  9. Blue-Shifted Green Fluorescent Protein Homologues Are Brighter than Enhanced Green Fluorescent Protein under Two-Photon Excitation

    Molina Rs, Tran Tm, Campbell Re, Lambert Gg, Salih A, Shaner Nc, Hughes Te, Drobizhev M

    (2017). The Journal of Physical Chemistry Letters, 8(12) , 2548-2554. doi: 10.1021/acs.jpclett.7b00960. Article   Pubmed

  10. Characterization of a spectrally diverse set of fluorescent proteins as FRET acceptors for mTurquoise2

    Mastop M, Bindels Ds, Shaner Nc, Postma M, Gadella Twj, Goedhart J

    (2017). Scientific Reports, 7(1) , 11999. doi: 10.1038/s41598-017-12212-x. Article   Pubmed

  11. Structural analysis of the bright monomeric yellow-green fluorescent protein mNeonGreen obtained by directed evolution

    Clavel D, Gotthard G, Von Stetten D, De Sanctis D, Pasquier H, Lambert Gg, Shaner Nc, Royant A

    (2016). Acta Crystallographica Section D Structural Biology, 72(12) , 1298-1307. doi: 10.1107/s2059798316018623. Article   Pubmed

  12. Comparative assessment of fluorescent proteins for in vivo imaging in an animal model system

    Heppert Jk, Dickinson Dj, Pani Am, Higgins Cd, Steward A, Ahringer J, Kuhn Jr, Goldstein B

    (2016). Molecular Biology of the Cell, 27(22) , 3385-3394. doi: 10.1091/mbc.e16-01-0063. Article   Pubmed

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