ArticleNucleic acids research2009
Sub-cellular trafficking and functionality of 2'-O-methyl and 2'-O-methyl-phosphorothioate molecular beacons.
Article in Nucleic acids research, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed.
- DNA/RNA Fluorescence Imaging by Synthetic Nucleic Acids.Advances in experimental medicine and biology · 2021Article
- Recent Advances in the Molecular Beacon Technology for Live-Cell Single-Molecule Imaging.iScience · 2020Review
- Intracellular Delivery by Membrane Disruption: Mechanisms, Strategies, and Concepts.Chemical reviews · 2018Review
- Engineering Novel Molecular Beacon Constructs to Study Intracellular RNA Dynamics and Localization.Genomics, proteomics & bioinformatics · 2017Review
- A molecular beacon-based approach for live-cell imaging of RNA transcripts with minimal target engineering at the single-molecule level.Scientific reports · 2017Article
- Monitoring integrity and localization of modified single-stranded RNA oligonucleotides using ultrasensitive fluorescence methods.PloS one · 2017Article
- 2'-O-methyl nucleotide modified DNA substrates influence the cleavage efficiencies of BamHI and BglII.Journal of biosciences · 2014Article
- Article
- Imaging the directed transport of single engineered RNA transcripts in real-time using ratiometric bimolecular beacons.PloS one · 2014Article
- Fluorescent probes for nucleic Acid visualization in fixed and live cells.Molecules (Basel, Switzerland) · 2013Review
- Quantitative assessment of ratiometric bimolecular beacons as a tool for imaging single engineered RNA transcripts and measuring gene expression in living cells.Nucleic acids research · 2013Article
- Article
- Effect of LNA- and OMeN-modified oligonucleotide probes on the stability and discrimination of mismatched base pairs of duplexes.Journal of biosciences · 2012Article
- Molecular beacons: powerful tools for imaging RNA in living cells.Journal of nucleic acids · 2011Article
- Intracellular traffic of oligodeoxynucleotides in and out of the nucleus: effect of exportins and DNA structure.Oligonucleotides · 2010Article
- Ratiometric bimolecular beacons for the sensitive detection of RNA in single living cells.Nucleic acids research · 2010Article
Corrections and comments
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Authors and funding
3 authors.
Funding
Abstract
Molecular beacons (MBs) have shown great potential for the imaging of RNAs within single living cells; however, the ability to perform accurate measurements of RNA expression can be hampered by false-positives resulting from nonspecific interactions and/or nuclease degradation. These false-positives could potentially be avoided by introducing chemically modified oligonucleotides into the MB design. In this study, fluorescence microscopy experiments were performed to elucidate the subcellular trafficking, false-positive signal generation, and functionality of 2'-O-methyl (2Me) and 2'-O-methyl-phosphorothioate (2MePS) MBs. The 2Me MBs exhibited rapid nuclear sequestration and a gradual increase in fluorescence over time, with nearly 50% of the MBs being opened nonspecifically within 24 h. In contrast, the 2MePS MBs elicited an instantaneous increase in false-positives, corresponding to approximately 5-10% of the MBs being open, but little increase was observed over the next 24 h. Moreover, trafficking to the nucleus was slower. After 24 h, both MBs were localized in the nucleus and lysosomal compartments, but only the 2MePS MBs were still functional. When the MBs were retained in the cytoplasm, via conjugation to NeutrAvidin, a significant reduction in false-positives and improvement in functionality was observed. Overall, these results have significant implications for the design and applications of MBs for intracellular RNA measurement.
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