![]() ![]() Principles of Fluorescence Spectroscopy, 452 (Klewer Academic/Plenum Publishers, 1999).Ĭhen, R. Using droplet-based microfluidics to improve the catalytic properties of RNA under multiple-turnover conditions. In vitro selection of RNA molecules that bind specific ligands. Systematic evolution of ligands by exponential enrichment - RNA ligands to bacteriophage-T4 DNA-polymerase. Shape complementarity at protein/protein interfaces. Mg 2+ ions: do they bind to nucleobase nitrogens? Nucleic Acids Res. Crystal structures of the Mango-II RNA aptamer reveal heterogeneous fluorophore binding and guide engineering of variants with improved selectivity and brightness. Advanced fluorescence microscopy techniques–FRAP, FLIP, FLAP, FRET and FLIM. Structural principles of fluorescent RNA aptamers. Thiazole orange-peptide conjugates: sensitivity of DNA binding to chemical structure. The interactions between the fluorescent dye thiazole orange and DNA. Structural basis for high-affinity fluorophore binding and activation by RNA Mango. The growing and glowing toolbox of fluorescent and photoactive proteins. Fluorogenic RNA Mango aptamers for imaging small non-coding RNAs in mammalian cells. Imaging RNA polymerase III transcription using a photostable RNA-fluorophore complex. RNA Mango aptamer-fluorophore: a bright, high-affinity complex for RNA labeling and tracking. Fluorescence imaging of cellular metabolites with RNA. The improved iMango-III and Mango-III(A10U) fluoresce ~50% brighter than enhanced green fluorescent protein, making them suitable tags for live cell RNA visualization. The fluorophore is restrained into a planar conformation by the G-quadruplex, a lone, long-range trans Watson–Crick pair (whose A10U mutation increases quantum yield to 0.66), and a pyrimidine perpendicular to the nucleobase planes of those motifs. The structures reveal a globular architecture arising from an unprecedented pseudoknot-like connectivity between a G-quadruplex and an embedded non-canonical duplex. We report crystal structures of TO1-Biotin complexes of Mango-III, a structure-guided mutant Mango-III(A10U), and a functionally reselected mutant iMango-III. Uniquely among related aptamers, Mango-III exhibits biphasic thermal melting, characteristic of molecules with tertiary structure. Among these, the ~30 nucleotide Mango-III is notable because it binds the thiazole orange derivative TO1-Biotin with high affinity and fluoresces brightly (quantum yield 0.55). Several turn-on RNA aptamers that activate small-molecule fluorophores have been selected in vitro. ![]()
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