scholarly journals Bright, fluorogenic and photostable avidity probes for RNA imaging

2021 ◽  
Author(s):  
Bastian B&uumlhler ◽  
Anja Benderoth ◽  
Daniel Englert ◽  
Franziska Gr&uumln ◽  
Janin Schokolowski ◽  
...  
Keyword(s):  
Dissociation Constants ◽  
Living Cells ◽  
Fluorescent Light ◽  
Binding Strength ◽  
Rna Imaging ◽  
High Fluorescence

Fluorescent light-up aptamers (FLAPs) emerged as valuable tools to visualize RNA, but are mostly limited by poor brightness, low photostability and high fluorescence background. In this study, we combine bivalent (silicon) rhodamine fluorophores with dimeric FLAPs to yield bright and photostable complexes with low picomolar dissociation constants. Our avidity-based approach resulted in extreme binding strength and high fluorogenicity, enabling single mRNA tracking in living cells.

scholarly journals Visualization of ATP levels inside single living cells with fluorescence resonance energy transfer-based genetically encoded indicators

2009 ◽  
Vol 106 (37) ◽  
pp. 15651-15656 ◽  
Author(s):  
Hiromi Imamura ◽  
Kim P. Huynh Nhat ◽  
Hiroko Togawa ◽  
Kenta Saito ◽  
Ryota Iino ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Atp Synthase ◽  
Hela Cells ◽  
Dissociation Constants ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Living Cells ◽  
Atp Levels ◽  
Fluorescence Resonance

Adenosine 5′-triphosphate (ATP) is the major energy currency of cells and is involved in many cellular processes. However, there is no method for real-time monitoring of ATP levels inside individual living cells. To visualize ATP levels, we generated a series of fluorescence resonance energy transfer (FRET)-based indicators for ATP that were composed of the ε subunit of the bacterial FoF1-ATP synthase sandwiched by the cyan- and yellow-fluorescent proteins. The indicators, named ATeams, had apparent dissociation constants for ATP ranging from 7.4 μM to 3.3 mM. By targeting ATeams to different subcellular compartments, we unexpectedly found that ATP levels in the mitochondrial matrix of HeLa cells are significantly lower than those of cytoplasm and nucleus. We also succeeded in measuring changes in the ATP level inside single HeLa cells after treatment with inhibitors of glycolysis and/or oxidative phosphorylation, revealing that glycolysis is the major ATP-generating pathway of the cells grown in glucose-rich medium. This was also confirmed by an experiment using oligomycin A, an inhibitor of FoF1-ATP synthase. In addition, it was demonstrated that HeLa cells change ATP-generating pathway in response to changes of nutrition in the environment.

A fluorescent light-up probe based on AIE and ESIPT processes for β-galactosidase activity detection and visualization in living cells

2015 ◽  
Vol 3 (47) ◽  
pp. 9168-9172 ◽  
Author(s):  
Lu Peng ◽  
Meng Gao ◽  
Xiaolei Cai ◽  
Ruoyu Zhang ◽  
Kai Li ◽  
...  
Keyword(s):  
Fluorescent Probe ◽  
Living Cells ◽  
Fluorescent Light ◽  
Galactosidase Activity ◽  
Activity Detection

A fluorescent probe with both AIE and ESIPT characteristics is developed for a β-galactosidase activity study in solution and living cells.

Genetically encoded tools for RNA imaging in living cells

2015 ◽  
Vol 31 ◽  
pp. 42-49 ◽  
Author(s):  
Anna K. Rath ◽  
Andrea Rentmeister
Keyword(s):  
Living Cells ◽  
Rna Imaging

scholarly journals Background free tracking of single RNA in living cells using catalytically inactive CasE

2019 ◽  
Author(s):  
Feng Gao ◽  
Yue Sun ◽  
Feng Jiang ◽  
Xiaoyue Bai ◽  
Chunyu Han
Keyword(s):  
Living Cells ◽  
The Other ◽  
Imaging Systems ◽  
Essential Information ◽  
Rna Imaging

AbstractRNAs have important and diverse functions. Visualizing an isolated RNA in living cells provide us essential information of its roles. By now, there are two kinds of live RNA imaging systems invented, one is the MS2 system and the other is the Cas13a system. In this study, we show that when fused with split-Fp, CasE can be engineered into a live RNA tracking tool.

scholarly journals Binding strength and activation state of single fibrinogen-integrin pairs on living cells

2002 ◽  
Vol 99 (11) ◽  
pp. 7426-7431 ◽  
Author(s):  
R. I. Litvinov ◽  
H. Shuman ◽  
J. S. Bennett ◽  
J. W. Weisel
Keyword(s):  
Living Cells ◽  
Binding Strength

scholarly journals Development of a red fluorescent light-up probe for highly selective and sensitive detection of vicinal dithiol-containing proteins in living cells

2016 ◽  
Vol 7 (1) ◽  
pp. 518-524 ◽  
Author(s):  
Yuanyuan Wang ◽  
Xiao-Feng Yang ◽  
Yaogang Zhong ◽  
Xueyun Gong ◽  
Zheng Li ◽  
...  
Keyword(s):  
Living Cells ◽  
Sensitive Detection ◽  
Fluorescent Light

An environment-sensitive red fluorescent light-up probe for vicinal dithiol-containing proteins (VDPs) in living cells has been successfully developed.

scholarly journals Current techniques for visualizing RNA in cells

F1000Research ◽  
2016 ◽  
Vol 5 ◽  
pp. 775 ◽  
Author(s):  
Lilith V.J.C. Mannack ◽  
Sebastian Eising ◽  
Andrea Rentmeister
Keyword(s):  
Binding Proteins ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
State Of The Art ◽  
Living Cells ◽  
Enzymatic Modification ◽  
Current State ◽  
Rna Imaging ◽  
In Cells ◽  
Art Techniques

Labeling RNA is of utmost interest, particularly in living cells, and thus RNA imaging is an emerging field. There are numerous methods relying on different concepts ranging from hybridization-based probes, over RNA-binding proteins to chemo-enzymatic modification of RNA. These methods have different benefits and limitations. This review aims to outline the current state-of-the-art techniques and point out their benefits and limitations.

scholarly journals Live-cell imaging of multiple endogenous mRNAs permits the direct observation of RNA granule dynamics

2018 ◽  
Vol 54 (52) ◽  
pp. 7151-7154 ◽  
Author(s):  
Kenji Yatsuzuka ◽  
Shin-ichi Sato ◽  
Kathleen Beverly Pe ◽  
Yousuke Katsuda ◽  
Ippei Takashima ◽  
...  
Keyword(s):  
Direct Observation ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Living Cells ◽  
Live Cell ◽  
Stress Granule ◽  
Granule Formation ◽  
Imaging Technology ◽  
Rna Imaging

A multicolor RNA imaging technology permits the direct observation of stress granule formation in living cells.

scholarly journals A simple sensor based on 1,8-naphthalimide with large Stokes shift for detection of hypochlorous acid in living cells

RSC Advances ◽  
2019 ◽  
Vol 9 (54) ◽  
pp. 31196-31201 ◽  
Author(s):  
Chunpeng Jiao ◽  
Yuanyuan Liu ◽  
Wenjuan Lu ◽  
Pingping Zhang ◽  
Xia Ma ◽  
...  
Keyword(s):  
Detection Limit ◽  
Quantum Yield ◽  
Fluorescence Quantum Yield ◽  
Hypochlorous Acid ◽  
Stokes Shift ◽  
High Sensitivity ◽  
Living Cells ◽  
High Fluorescence ◽  
Excellent Selectivity ◽  
High Fluorescence Quantum Yield

Probe 1 shows excellent selectivity to HOCl among different ions including common ROS, high sensitivity, high fluorescence quantum yield (Φ = 0.93), low detection limit (0.237 μM) and successfully used for the detection of HOCl in cells.

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