Delivery of molecular beacons for live-cell... [Methods Mol Biol. 2011. Www.springerlink.com/content/l439737l8l454r0v/fulltext.pdf. Over the past decade, a variety of oligonucleotide-based probes have been developed that allow for direct visualization of RNA molecules in living cells.
Of these, molecular beacons have garnered a particularly high degree of interest due to their simple yet exquisite unimolecular stem-loop design that allows for the efficient conversion of target recognition into a specific fluorescent signal. As a result of their favorable fluorescent enhancement and their high specificity, molecular beacons have been used for a wide range of applications, including the monitoring of RNA expression and localization in living cells, cancer cell detection, and the study of viral infections.
In this chapter we describe a general methodology that can be followed for the imaging and analysis of RNA in living cells using molecular beacons. Modification Highlight: Photo-Cleavable Spacer. Photo-cleavable (PC) modifications contain a photolabile functional group that is cleavable by UV light of specific wavelength (300–350 nm).
The PC spacer is a 10-atom linker arm that can only be cleaved when exposed to UV light within the appropriate spectral range. The resulting oligo will have a 5’ phosphate group that is available for subsequent ligase reactions. PC spacers can be placed between DNA bases or between an oligo and a terminal modification such as a fluorophore. Quenched probes for highly specific detect... [Trends Biotechnol. 2005. Ratiometric bimolecular beacons for the se... [Nucleic Acids Res. 2010. Efficient cytosolic delivery of molecular ... [Nucleic Acids Res. 2008.
Avoiding false-positive signals with nucle... [Nucleic Acids Res. 2007. Hybridization kinetics and thermodynamics ... [Nucleic Acids Res. 2003. Spectroscopic features of dual fluorescence/lumine... [Anal Chem. 2003. Molecular beacons: an optimal mul... [Biochem Biophys Res Commun. 2008. Sub-cellular trafficking and functionality... [Nucleic Acids Res. 2009.
Structure–function relationships of shared-stem and conventional molecular beacons. Shedding light on health and d... [Brief Funct Genomic Proteomic. 2003. Molecular beacons: a novel DNA probe for nucleic a... [Chemistry. 2000.