TO1-3PEG-Desthiobiotin Fluorophore

Cat. No.
G7956
Unit
250 µM (100 µl)
Price
$375.00
Cat. No. G7956
Name TO1-3PEG-Desthiobiotin Fluorophore
Unit 250 µM (100 µl)
Category RNA Tracking (RNA Mango)
Description

RNA Mango technology is based on the specific binding of the RNA Mango Aptamer and a Thizole Orange (TO) bi-functional dye. The main features of this technology is the tight binding between the dye and aptamer (KD ≈ 3nM) , and the strong ~1000X enhancement of the dye’s fluorescence when bound to the Mango apatmer (Fluorescent enhancement FE=1,100).
The TO dye has a number of other desirable properties including:

  • small size
  • lack of toxicity
  • plasma and nuclear membrane permeability
  • short intracellular half-life
  • the accessibility of a broad wavelength range simply via substitutions and alterations to the TO structure

TO1-biotin is the standard variety of TO dye for RNA Mango experiments; other dye variants are available. Learn more about the RNA Mango technology here.

 

Watch RNA Mango in Action

Transcription reaction were carried out in 300 µL volumes using T7 RNA polymerase (400 U, 50U/µL, applied biological materials), 0.5 µM TO1-3PEG-Biotin (applied biological materials), in 8 mM GTP, 5 mM CTP and ATP, 2 mM UTP, 40 mM TRIS buffer pH 7.9, 2.5 mM spermidine, 26 mM MgCl2, 20 mM KCl, Pyrophosphatase (0.5 U, 0.1 U/µL, ThermoFisher Scientific), and 0.01% Triton X-100. To each sample, either water (Negative), 0.33 µM DNA template (Mango Transcription), or 500 nM final Mango III A10U RNA (Positive) was added. Samples were visualized in a blue light box, movie is played back at 30X speed.

 

The most advanced RNA tracking, visualization and pull down technology.

Technology for studying the diverse cellular roles of RNA has lagged behind the tools for studying DNA and proteins, but innovative researchers are working to change that! One such researcher is Dr. Peter Unrau of Simon Fraser University. He and his team have created RNA Mango, a novel technology with a number of useful applications.

Storage Condition Stored at -20°C. Protect from light.
Depositor Unrau Lab
Material Citation If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. G7956
Datasheet
Search CoA here
MSDS
Supporting Protocol
  • Trachman RJ 3rd, Demeshkina NA, Lau MWL, Panchapakesan SSS, Jeng SCY, Unrau PJ, Ferré-D'Amaré AR. et al. "Structural basis for high-affinity fluorophore binding and activation by RNA Mango." Nat Chem Biol. 2017 Jul;13(7):807-813. DOI: 10.1038/nchembio.2392.
  • Jeng SC, Chan HH, Booy EP, McKenna SA, Unrau PJ. et al. "Fluorophore ligand binding and complex stabilization of the RNA Mango and RNA Spinach aptamers." RNA. 2016 Dec;22(12):1884-1892. Epub 2016 Oct 24. PubMed : 27777365.
  • Ouellet J. et al. "RNA Fluorescence with Light-Up Aptamers." Front Chem. 2016 Jun 28;4:29. DOI: 10.3389/fchem.2016.00029. eCollection 2016. Review. PubMed : 27446908.
  • Panchapakesan SS, Jeng SC, Unrau PJ. et al. "RNA complex purification using high-affinity fluorescent RNA aptamer tags." Ann N Y Acad Sci. 2015 Apr;1341:149-55. DOI: 10.1111/nyas.12663. Epub 2015 Jan 13.
  • Dolgosheina EV, Jeng SC, Panchapakesan SS, Cojocaru R, Chen PS, Wilson PD, Hawkins N, Wiggins PA, Unrau PJ. et al. "RNA mango aptamer-fluorophore: a bright, high-affinity complex for RNA labeling and tracking." ACS Chem Biol. 2014 Oct 17;9(10):2412-20. DOI: 10.1021/cb500499x. Epub 2014 Aug 21.
  • Autour, A., C Y Jeng, S., D Cawte, A., Abdolahzadeh, A., Galli, A., Panchapakesan, S. S. S., Rueda, D., Ryckelynck, M., & Unrau, P. J. (2018). Fluorogenic RNA Mango aptamers for imaging small non-coding RNAs in mammalian cells. Nature communications9(1), 656. https://doi.org/10.1038/s41467-018-02993-8 
  • Trachman, R. J., & Ferré-D'Amaré, A. R. (2019). Tracking RNA with light: selection, structure, and design of fluorescence turn-on RNA aptamers. Quarterly reviews of biophysics52, e8. https://doi.org/10.1017/S0033583519000064 
  • Trachman, R. J., 3rd, Autour, A., Jeng, S. C. Y., Abdolahzadeh, A., Andreoni, A., Cojocaru, R., Garipov, R., Dolgosheina, E. V., Knutson, J. R., Ryckelynck, M., Unrau, P. J., & Ferré-D'Amaré, A. R. (2019). Structure and functional reselection of the Mango-III fluorogenic RNA aptamer. Nature chemical biology15(5), 472–479. https://doi.org/10.1038/s41589-019-0267-9 
  • Kong, K. Y. S., Jeng, S. C. Y., Rayyan, B., & Unrau, P. J. (2021). RNA Peach and Mango: Orthogonal two-color fluorogenic aptamers distinguish nearly identical ligands. RNA (New York, N.Y.)27(5), 604–615. Advance online publication. https://doi.org/10.1261/rna.078493.120 
  • Cawte, A. D., Unrau, P. J., & Rueda, D. S. (2020). Live cell imaging of single RNA molecules with fluorogenic Mango II arrays. Nature communications11(1), 1283. https://doi.org/10.1038/s41467-020-14932-7 
  • Panchapakesan, S. S. S., Ferguson, M. L., Hayden, E. J., Chen, X., Hoskins, A. A., & Unrau, P. J. (2017). Ribonucleoprotein purification and characterization using RNA Mango. RNA (New York, N.Y.)23(10), 1592–1599. https://doi.org/10.1261/rna.062166.117 
  • Mitra, J., & Ha, T. (2019). Nanomechanics and co-transcriptional folding of Spinach and Mango. Nature communications10(1), 4318. https://doi.org/10.1038/s41467-019-12299-y 
  • Shi, J., Gao, X., Tian, T., Yu, Z., Gao, B., Wen, A., You, L., Chang, S., Zhang, X., Zhang, Y., & Feng, Y. (2019). Structural basis of Q-dependent transcription antitermination. Nature communications10(1), 2925. https://doi.org/10.1038/s41467-019-10958-8 
  • Fang, C., Philips, S. J., Wu, X., Chen, K., Shi, J., Shen, L., Xu, J., Feng, Y., O'Halloran, T. V., & Zhang, Y. (2021). CueR activates transcription through a DNA distortion mechanism. Nature chemical biology17(1), 57–64. https://doi.org/10.1038/s41589-020-00653-x 
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