scholarly journals Labeling RNAs in Live Cells Using Malachite Green Aptamer Scaffolds as Fluorescent Probes

2018 ◽  
Vol 7 (3) ◽  
pp. 758-766 ◽  
Author(s):  
V. Siddartha Yerramilli ◽  
Kyung Hyuk Kim
Keyword(s):  
Malachite Green ◽  
Fluorescent Probes ◽  
Live Cells ◽  
Malachite Green Aptamer

scholarly journals Labeling RNAs in live cells using malachite green aptamer scaffolds as fluorescent probes

2017 ◽  
Author(s):  
V. Siddartha Yerramilli ◽  
Kyung Hyuk Kim
Keyword(s):  
Malachite Green ◽  
Tandem Repeats ◽  
Fluorescent Proteins ◽  
Rna Aptamers ◽  
Live Cells ◽  
High Background ◽  
Rna Molecules ◽  
High Background Noise ◽  
Fluorescent Tags ◽  
Malachite Green Aptamer

AbstractRNAs mediate many different processes that are central to cellular function. The ability to quantify or image RNAs in live cells is very useful in elucidating such functions of RNA. RNA aptamerfluorogen systems have been increasingly used in labeling RNAs in live cells. Here, we use the malachite green aptamer (MGA), an RNA aptamer that can specifically bind to malachite green (MG) dye and induces it to emit far-red fluorescence signals. Previous studies on MGA showed a potential for the use of MGA for genetically tagging other RNA molecules in live cells. However, these studies also exhibited low fluorescence signals and high background noise. Here we constructed and tested RNA scaffolds containing multiple tandem repeats of MGA as a strategy to increase the brightness of the MGA aptamer-fluorogen system as well as to make the system fluoresce when tagging various RNA molecules, in live cells. We demonstrate that our MGA scaffolds can induce fluorescence signals by up to ~20 fold compared to the basal level as a genetic tag for other RNA molecules. We also show that our scaffolds function reliably as genetically-encoded fluorescent tags for mRNAs of fluorescent proteins and other RNA aptamers.

scholarly journals Fluorescent Probes for Live Cell Thiol Detection

Molecules ◽  
2021 ◽  
Vol 26 (12) ◽  
pp. 3575
Author(s):  
Shenggang Wang ◽  
Yue Huang ◽  
Xiangming Guan
Keyword(s):  
Fluorescent Probes ◽  
Biological System ◽  
High Sensitivity ◽  
Intracellular Distribution ◽  
Live Cell ◽  
Live Cells ◽  
High Demand ◽  
Non Invasive

Thiols play vital and irreplaceable roles in the biological system. Abnormality of thiol levels has been linked with various diseases and biological disorders. Thiols are known to distribute unevenly and change dynamically in the biological system. Methods that can determine thiols’ concentration and distribution in live cells are in high demand. In the last two decades, fluorescent probes have emerged as a powerful tool for achieving that goal for the simplicity, high sensitivity, and capability of visualizing the analytes in live cells in a non-invasive way. They also enable the determination of intracellular distribution and dynamitic movement of thiols in the intact native environments. This review focuses on some of the major strategies/mechanisms being used for detecting GSH, Cys/Hcy, and other thiols in live cells via fluorescent probes, and how they are applied at the cellular and subcellular levels. The sensing mechanisms (for GSH and Cys/Hcy) and bio-applications of the probes are illustrated followed by a summary of probes for selectively detecting cellular and subcellular thiols.

scholarly journals Fluorescent Probes with Unnatural Amino Acids to Monitor Proteasome Activity in Real-Time

2020 ◽  
Author(s):  
Breanna L. Zerfas ◽  
Rachel A. Coleman ◽  
Andres Salazar Chaparro ◽  
Nathaniel J. Macatangay ◽  
Darci Trader
Keyword(s):  
Amino Acids ◽  
Small Molecule ◽  
Fluorescent Probes ◽  
Unnatural Amino Acids ◽  
Cell Types ◽  
Proteasome Activity ◽  
Live Cells ◽  
Fluorescent Substrate ◽  
The Individual ◽  
Small Molecule Modulators

<div> <div> <div> <p>The proteasome is an essential protein complex that, when dysregulated, can result in various diseases in eukaryotic cells. As such, understanding the enzymatic activity of the proteasome and what can alter it is crucial to elucidating its roles in these diseases. This can be done effectively by using activity-based fluorescent substrate probes, of which there are many commercially available that target the individual protease-like subunits in the 20S CP of the proteasome. Unfortunately, these probes have not displayed appropriate characteristics for their use in live cell-based assays. In the work presented here, we have developed a set of probes which have shown improved fluorescence properties and selectivity towards the proteasome compared to other cellular proteases. By including unnatural amino acids, we have found probes which can be utilized in various applications, including monitoring the effects of small molecule stimulators of the proteasome in live cells and comparing the relative proteasome activity across different cancer cell types. In future studies, we expect the fluorescent probes presented here will serve as tools to support the discovery and characterization of small molecule modulators of proteasome activity. </p> </div> </div> </div>

Harnessing Se=N to develop novel fluorescent probes for visualizing the variation of endogenous hypobromous acid (HOBr) during the administration of immunotherapeutic agent

2021 ◽  
Author(s):  
Jian Zhang ◽  
kaiqiang Liu ◽  
Jingwen Li ◽  
Yingying Xie ◽  
Yong Li ◽  
...  
Keyword(s):  
Fluorescent Probes ◽  
Live Cells ◽  
Immunotherapeutic Agent ◽  
Hypobromous Acid ◽  
In Virtue Of

In virtue of the formation of Se=N, ABT-Se and NDI-Se were developed to detect and visualize endogenous HOBr in live cells. Specifically, the upregulation of HOBr was monitored by NDI-Se...

Turn-on visible and ratiometric near-infrared fluorescent probes for distinction endogenous esterases and chymotrypsins in live cells

2020 ◽  
Vol 306 ◽  
pp. 127567 ◽  
Author(s):  
Jiemin Wang ◽  
Zhidong Teng ◽  
Ting Cao ◽  
Jing Qian ◽  
Lei Zheng ◽  
...  
Keyword(s):  
Fluorescent Probes ◽  
Near Infrared ◽  
Live Cells ◽  
Turn On

scholarly journals A dual-labeling probe to track functional mitochondria–lysosome interactions in live cells

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Qixin Chen ◽  
Hongbao Fang ◽  
Xintian Shao ◽  
Zhiqi Tian ◽  
Shanshan Geng ◽  
...  
Keyword(s):  
Fluorescence Intensity ◽  
Fluorescent Probes ◽  
Living Cells ◽  
Molecular Probe ◽  
Live Cells ◽  
Blue Fluorescence ◽  
Red Fluorescence ◽  
Dynamic Tracking ◽  
A Cell

AbstractMitochondria–lysosome interactions are essential for maintaining intracellular homeostasis. Although various fluorescent probes have been developed to visualize such interactions, they remain unable to label mitochondria and lysosomes simultaneously and dynamically track their interaction. Here, we introduce a cell-permeable, biocompatible, viscosity-responsive, small organic molecular probe, Coupa, to monitor the interaction of mitochondria and lysosomes in living cells. Through a functional fluorescence conversion, Coupa can simultaneously label mitochondria with blue fluorescence and lysosomes with red fluorescence, and the correlation between the red–blue fluorescence intensity indicates the progress of mitochondria–lysosome interplay during mitophagy. Moreover, because its fluorescence is sensitive to viscosity, Coupa allowed us to precisely localize sites of mitochondria–lysosome contact and reveal increases in local viscosity on mitochondria associated with mitochondria–lysosome contact. Thus, our probe represents an attractive tool for the localization and dynamic tracking of functional mitochondria–lysosome interactions in living cells.

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