Cell chemical biology, Journal Year: 2020, Volume and Issue: 27(8), P. 921 - 936
Published: July 30, 2020
Language: Английский
Journal of the American Chemical Society, Journal Year: 2019, Volume and Issue: 141(18), P. 7562 - 7571
Published: April 15, 2019
Although genetically encoded light-up RNA aptamers have become promising tools for visualizing and tracking RNAs in living cells, aptamer/ligand pairs that emit the far-red near-infrared (NIR) regions are still rare. In this work, we developed a aptamer binds silicon rhodamines (SiRs). SiRs photostable, NIR-emitting fluorophores change their open–closed equilibrium between noncolored spirolactone fluorescent zwitterion response to environment. This property is responsible high cell permeability fluorogenic behavior. Aptamers binding SiR were vitro selected from combinatorial library. Sequencing, bioinformatic analysis, truncation, mutational studies revealed 50-nucleotide minimal aptamer, SiRA, which with nanomolar affinity target SiR. addition rhodamines, SiRA structurally related carborhodamines, making it versatile tool spanning region of spectrum. Photophysical characterization showed remarkably resistant photobleaching constitutes brightest system known date owing its favorable features: fluorescence quantum yield 0.98 an extinction coefficient 86 000 M–1cm–1. Using system, visualized expression bacteria no-wash live-cell imaging experiments also report stimulated emission depletion (STED) super-resolution microscopy images aptamer-based, fluorescently labeled mRNA live cells. work represents, our knowledge, first application popular dyes intramolecular spirocyclization as means background reduction field aptamer-based imaging. We anticipate potential novel labeling address biological questions.
Language: Английский
Citations
125
Annual Review of Biophysics, Journal Year: 2018, Volume and Issue: 47(1), P. 85 - 106
Published: Jan. 18, 2018
RNA is the fundamental information transfer system in cell. The ability to follow single messenger RNAs (mRNAs) from transcription degradation with fluorescent probes gives quantitative about how transferred DNA proteins. This review focuses on latest technological developments field of single-mRNA detection and their usage study gene expression both fixed live cells. By describing application these imaging tools, we journey mRNA decay cells, single-molecule resolution. We current theoretical models for translation that were generated by single-cell studies. These methods provide a basis interactions generate phenotypes, fundamentally changing our understating regulation.
Language: Английский
Citations
122
Nature Communications, Journal Year: 2020, Volume and Issue: 11(1)
Published: March 9, 2020
RNA molecules play vital roles in many cellular processes. Visualising their dynamics live cells at single-molecule resolution is essential to elucidate role metabolism. aptamers, such as Spinach and Mango, have recently emerged a powerful background-free technology for live-cell imaging due fluorogenic properties upon ligand binding. Here, we report novel array of Mango II aptamers fixed with high contrast sensitivity. Direct comparison MS2-tdMCP-mCherry dual-labelled mRNAs show marked improvements signal noise ratio using the aptamers. Using both coding (β-actin mRNA) long non-coding (NEAT1) RNAs, that does not affect localisation. Additionally, can track single extended time periods, likely bleached fluorophore replacement. This property makes arrays readily compatible structured illumination super-resolution microscopy.
Language: Английский
Citations
120
Nature Structural & Molecular Biology, Journal Year: 2018, Volume and Issue: 25(12), P. 1077 - 1085
Published: Nov. 23, 2018
Language: Английский
Citations
119
Molecular Cell, Journal Year: 2019, Volume and Issue: 74(3), P. 521 - 533.e6
Published: April 2, 2019
Language: Английский
Citations
113
Annual Review of Biochemistry, Journal Year: 2018, Volume and Issue: 88(1), P. 635 - 659
Published: Oct. 25, 2018
In the past decades, advances in microscopy have made it possible to study dynamics of individual biomolecules vitro and resolve intramolecular kinetics that would otherwise be hidden ensemble averages. More recently, single-molecule methods been used image, localize, track individually labeled macromolecules cytoplasm living cells, allowing investigations intermolecular under physiologically relevant conditions. this review, we illuminate particular advantages techniques when studying cells discuss solutions specific challenges associated with these methods.
Language: Английский
Citations
112
Journal of Molecular Biology, Journal Year: 2018, Volume and Issue: 430(23), P. 4685 - 4701
Published: May 10, 2018
Language: Английский
Citations
110
Nature Chemical Biology, Journal Year: 2019, Volume and Issue: 16(1), P. 69 - 76
Published: Oct. 21, 2019
Language: Английский
Citations
108
Cell chemical biology, Journal Year: 2020, Volume and Issue: 27(8), P. 891 - 903
Published: July 7, 2020
Language: Английский
Citations
92
Angewandte Chemie International Edition, Journal Year: 2019, Volume and Issue: 59(11), P. 4511 - 4518
Published: Dec. 18, 2019
Abstract Spinach and Broccoli are fluorogenic RNA aptamers that bind DFHBI, a mimic of the chromophore in green fluorescent protein, activate its fluorescence. Spinach/Broccoli‐DFHBI complexes exhibit high fluorescence vitro, but they lower mammalian cells. Here, computational screening was used to identify BI, DFHBI derivative binds with higher affinity leads markedly cells compared previous ligands. BI prevents thermal unfolding at 37 °C, leading more folded thus Broccoli‐BI photostable owing impaired photoisomerization rapid unbinding photoisomerized cis ‐BI. These properties enable single mRNA containing 24 be imaged live treated BI. Small molecule ligands can promote folding cells, allow imaging aptamers.
Language: Английский
Citations
87