RNA Tracking & Visualization (RNA Mango)

RNA Tracking & Visualization (RNA Mango)

HOME abm Molecular Biology and PCR RNA Tracking & Visualization (RNA Mango)

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.

Key Features

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 aptamer (Fluorescent enhancement FE=1,100).

The Thizole Orange (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
The TO1 and TO3 dyes can be used in a 2-color reporter assay system using both the RNA Mango and RNA Peach aptamer systems (Kong et. al. 2021)

TO1-biotin is the standard variety of TO dye for RNA Mango experiments. View our complete list of RNA Mango dyes.

Can’t find what you’re looking for, or have a specific request?
Speak to one of our technical support specialists today technical@abmgood.com.

Popular Products

Documents

The RNA Mango Workflow

The RNA Mango Aptamer
The system has two components: the RNA Mango aptamer, and the TO-1 dye. The dye only fluoresces when bound to the Mango aptamer.

Tagging Your RNA
For mRNA inset the tag into the 3’UTR. For structured non-coding RNA replace a stem-loop that is not essential to the RNA function.

Application: RNA Visualization
Express RNA of interest (with Mango Tag) using a vector or CRISPR Knock-In. Soak cells in TO-1 Biotin dye to illuminate RNA of interest localization

Application: RNA-RNA or RNA-protein complex pulldown
Express RNA of interest (with Mango Tag) using a vector or CRISPR Knock-In. Lyse the cells, and recover RNA of interest with bound RNA or proteins using streptavidin breads

Laboratory Results

Yeast U1 Ribonucleoprotein (RNP) Complex Pulldown Purification
Left: RNAs present in native extract in which U1M migrates as doublet. Right: U1M is a single band RNP present after mango-based purification using TO1 3PEG-Desthiobiotin Fluorophore

RNA Mango in a Test Tube
Mango dye binding to in vitro transcribed RNA Mango-tagged sgRNA. Stable binding and fluorescence even after leaving the tube for 1 month at room temperature.

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.

RNA Mango Binding, Fluorescence, and Resistance Properties
Includes extensive citations list.

TO1 and TO3 dyes can be used in a 2-color reporter assay system using both the RNA Mango and RNA Peach aptamer systems (Kong et. al. 2021)

PRODUCT	UNIT	CAT.NO.
TO1-3PEG-Biotin Fluorophore	0.5 mg/ml (500 µl)	G955
TO1-3PEG-Desthiobiotin Fluorophore	0.5 mg/ml (500 µl)	G956
TO3-3PEG-Biotin Fluorophore	0.5 mg/ml (500 µl)	G959
YO3-3PEG-Biotin Fluorophore	0.5 mg/ml (500 µl)	G957