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.
RNA Mango is an aptamer–fluorophore system that can be used to visualize tagged RNA or recover the tagged RNA together with associated RNA and proteins.
The RNA Mango aptamer binds a Thiazole Orange (TO) bi-functional dye with high affinity, strongly activating fluorescence while retaining a biotin-based handle for affinity capture.
RNA Mango technology is based on the specific binding of the RNA Mango aptamer and a Thiazole Orange (TO) bi-functional dye. The main features are tight binding between the dye and aptamer (KD ≈ 3 nM) and an approximately 1,000-fold enhancement of the dye’s fluorescence when bound to the Mango aptamer (fluorescence enhancement FE = 1,100).
TO1-biotin is the standard variety of TO dye for RNA Mango experiments.
Choose a TO1, TO3, or YO3 fluorophore for RNA Mango imaging, reporter, and affinity-capture workflows.
| Product Name | Cat. No. | Size | Price |
|---|---|---|---|
| TO1-3PEG-Biotin Fluorophore | G7955 | 250 µM (100 µl) | |
| TO1-3PEG-Desthiobiotin Fluorophore | G7956 | 250 µM (100 µl) | |
| TO3-3PEG-Biotin Fluorophore | G7959 | 250 µM (100 µl) | |
| YO3-3PEG-Biotin Fluorophore | G7957 | 250 µM (100 µl) |
Follow the platform from aptamer tagging to RNA visualization or RNA–RNA and RNA–protein complex pull-down.
The system has two components: the RNA Mango aptamer and the TO-1 dye. The dye only fluoresces when bound to the Mango aptamer.
For mRNA, insert the tag into the 3′ UTR. For structured non-coding RNA, replace a stem-loop that is not essential to the RNA function.
Express RNA of interest with a Mango tag using a vector or CRISPR knock-in. Soak cells in TO-1 Biotin dye to illuminate the localization of the RNA of interest.
Express RNA of interest with a Mango tag using a vector or CRISPR knock-in. Lyse the cells, then recover the RNA of interest with bound RNA or proteins using streptavidin beads.
Review examples of RNP purification, RNA Mango fluorescence, long-term signal stability, transcription monitoring, and two-colour RNA Mango/RNA Peach reporter assays.
Left: RNAs present in native extract in which U1M migrates as a doublet. Right: U1M is a single-band RNP present after Mango-based purification using TO1-3PEG-Desthiobiotin Fluorophore.
Mango dye binding to in vitro transcribed RNA Mango-tagged sgRNA. Stable binding and fluorescence were observed even after leaving the tube for one month at room temperature.
Samples were visualized in a blue-light box, and the movie is played back at 30× speed.
Transcription reactions were carried out in 300 µL volumes using T7 RNA polymerase (400 U, 50 U/µL, applied biological materials), 0.5 µM TO1-3PEG-Biotin (applied biological materials), 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, Thermo Fisher 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.
Includes an extensive citations list.
TO1 and TO3 dyes can be used in a two-colour reporter assay system using both the RNA Mango and RNA Peach aptamer systems (Kong et al. 2021).
Use the original RNA Mango design and performance documents to plan aptamer placement, select dyes, and review reported binding, fluorescence, and resistance properties.
Guidance for placing the RNA Mango tag in mRNA and structured non-coding RNA targets.
Performance information for RNA Mango aptamer systems, including supporting references.
Browse the complete list of RNA Mango dyes, or speak with an abm technical support specialist at technical@abmgood.com.
Primary literature describing RNA Mango-based ribonucleoprotein purification, high-affinity fluorophore binding, and aptamer complex stabilization.
Concise answers to common questions about RNA Mango imaging, aptamer insertion, dye selection, and pull-down applications.
Connect with abm technical support for guidance on fluorophore selection, aptamer placement, RNA visualization, or pull-down workflow planning.
