ArticleNucleic acids research2008
Efficient cytosolic delivery of molecular beacon conjugates and flow cytometric analysis of target RNA.
Article in Nucleic acids research, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 41 papers.
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Who cites it
41 citing papers in PubMed.
- Delivering Molecular Beacons via an Electroporation-Based Approach Enables Live-Cell Imaging of Single RNA Transcripts and Genomic Loci.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Spuriously transcribed RNAs from CRISPR-sgRNA expression plasmids scaffold biomolecular condensate formation and hamper accurate genomic imaging.Nucleic acids research · 2025Article
- Molecular Dynamics Simulations of RNA Motifs to Guide the Architectural Parameters and Design Principles of RNA Nanostructures.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Complexation design of cationized gelatin and molecular beacon to visualize intracellular mRNA.PloS one · 2021Article
- Recent Advances in the Molecular Beacon Technology for Live-Cell Single-Molecule Imaging.iScience · 2020Review
- Nucleic-Acid Structures as Intracellular Probes for Live Cells.Advanced materials (Deerfield Beach, Fla.) · 2020Review
- Bioorthogonal mRNA labeling at the poly(A) tail for imaging localization and dynamics in live zebrafish embryos.Chemical science · 2020Article
- CRISPR/dual-FRET molecular beacon for sensitive live-cell imaging of non-repetitive genomic loci.Nucleic acids research · 2019Article
- Multiple covalent fluorescence labeling of eukaryotic mRNA at the poly(A) tail enhances translation and can be performed in living cells.Nucleic acids research · 2019Article
- Comparing Agent-Based Delivery of DNA and PNA Forced Intercalation (FIT) Probes for Multicolor mRNA Imaging.Chembiochem : a European journal of chemical biology · 2019Article
- Nucleic Acid Strand Displacement with Synthetic mRNA Inputs in Living Mammalian Cells.ACS synthetic biology · 2018Article
- A molecular beacon-based approach for live-cell imaging of RNA transcripts with minimal target engineering at the single-molecule level.Scientific reports · 2017Article
- Advancement of the Emerging Field of RNA Nanotechnology.ACS nano · 2017Review
- Protecting microRNAs from RNase degradation with steric DNA nanostructures.Chemical science · 2017Article
- Can we observe changes in mRNA "state"? Overview of methods to study mRNA interactions with regulatory proteins relevant in cancer related processes.The Analyst · 2016Review
- Barriers to Liposomal Gene Delivery: from Application Site to the Target.Iranian journal of pharmaceutical research : IJPR · 2016Review
- Beyond the Barriers of Light Penetration: Strategies, Perspectives and Possibilities for Photodynamic Therapy.Theranostics · 2016Review
- Non-genetic Purification of Ventricular Cardiomyocytes from Differentiating Embryonic Stem Cells through Molecular Beacons Targeting IRX-4.Stem cell reports · 2015Article
- Single-cell detection of mRNA expression using nanofountain-probe electroporated molecular beacons.Small (Weinheim an der Bergstrasse, Germany) · 2015Article
- Molecular beacon-enabled purification of living cells by targeting cell type-specific mRNAs.Nature protocols · 2014Article
Corrections and comments
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Authors and funding
3 authors.
Funding
Abstract
Fluorescent microscopy experiments show that when 2'-O-methyl-modified molecular beacons (MBs) are introduced into NIH/3T3 cells, they elicit a nonspecific signal in the nucleus. This false-positive signal can be avoided by conjugating MBs to macromolecules (e.g. NeutrAvidin) that prevent nuclear sequestration, but the presence of a macromolecule makes efficient cytosolic delivery of these probes challenging. In this study, we explored various methods including TAT peptide, Streptolysin O and microporation for delivering NeutrAvidin-conjugates into the cytosol of living cells. Surprisingly, all of these strategies led to entrapment of the conjugates within lysosomes within 24 h. When the conjugates were pegylated, to help prevent intracellular recognition, only microporation led to a uniform cytosolic distribution. Microporation also yielded a transfection efficiency of 93% and an average viability of 86%. When cells microporated with MB-NeutrAvidin conjugates were examined via flow cytometry, the signal-to-background was found to be more than 3 times higher and the sensitivity nearly five times higher than unconjugated MBs. Overall, the present study introduces an improved methodology for the high-throughput detection of RNA at the single cell level.
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