Molecular beacons and related probes for intracellular RNA imaging

Advanced Review

Philip J. Santangelo

Published Online: Sep 03 2009

DOI: 10.1002/wnan.52

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Accurately imaging endogenous or non‐engineered RNA in live cells is not an easy task. Ideally, a probe and imaging strategy will have the following properties: (1) functional probes will be delivered to the desired cellular compartment, (2) they will achieve the correct level of affinity to bind target RNA efficiently but not inhibit their function, (3) be sensitive enough to allow for the accurate detection of the cellular RNA population, and (4) allow for the tracking of RNA through biogenesis, transport, translation, and degradation pathways. In this review, the capabilities of current nucleic acid‐based probes and strategies used to image native RNA are discussed and analyzed, and probe and strategy recommendations for new users are given. The review is concluded by addressing topics for future research, all in the hope of achieving the ideal RNA imaging probe and strategy. WIREs Nanomed Nanobiotechnol 2010 2 11–19

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Figure 1.

Depiction of the two primary nucleic acid structures utilized for RNA imaging. (a) Linear nucleic acid with a single fluorescent label, and a depiction of the imaging strategy used for identifying bound probes. (b) Schematic of molecular beacon and its conversion from a dark to bright state upon hybridization and (c) depiction of how molecular beacons are used for fluorescence energy resonance transfer (FRET) imaging. (Reprinted with permission from Ref 17. Copyright 2004 Oxford University Press).

[ Normal View 54K | Magnified View 95K ]

Figure 2.

Imaging of β-actin mRNA colocalization with ZBP1 in a motile A549 cell; white arrow denotes lamellipodia. Plot represents pixel intensities along yellow line in image.

[ Normal View 43K | Magnified View 70K ]

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