A far red emissive RNA aptamer–fluorophore system for demethylase FTO detection: design and optimization

Gao, Wenjuan; Xu, Liren; Jing, Jing; Zhang, Xiaoling

doi:10.1039/d3nj00043e

New J. Chem.

2023

DOI: 10.1039/d3nj00043e

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A far red emissive RNA aptamer–fluorophore system for demethylase FTO detection: design and optimization

Wenjuan Gao

Liren Xu

Jing Jing

et al.

Abstract: Fat mass and obesity-associated protein (FTO) is a demethylase that participates in N6-methyladenosine (m6A) RNA modification. This demethylase previously linked to obesity and diabetes, is dysregulated in a number of...

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A Demethylation-Switchable Aptamer Design Enables Lag-Free Monitoring of m⁶A Demethylase FTO with Energy Self-Sufficient and Structurally Integrated Features

Shi,

Lei,

Chen

et al. 2024

J. Am. Chem. Soc.

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Cellular context profiling of modification effector proteins is critical for an in-depth understanding of their biological roles in RNA N 6 -methyladenosine (m 6 A) modification regulation and function. However, challenges still remain due to the high context complexities, which call for a versatile toolbox for accurate live-cell monitoring of effectors. Here, we propose a demethylationswitchable aptamer sensor engineered with a site-specific m 6 A (DSA-m 6 A) for lag-free monitoring of the m 6 A demethylase FTO activity in living cells. As a proof of concept, a DNA aptamer against adenosine triphosphate (ATP) is selected to construct the DSA-m 6 A model, as the "universal energy currency" role of ATP could guarantee the equally fast and spontaneous conformation change of DSA-m 6 A sensor upon demethylation and ATP binding in living organisms, thus enabling sensitive monitoring of FTO activity with neither time delay nor recourse to extra supply of substances. This ATP-driven DSA-m 6 A design facilitates biomedical research, including live-cell imaging, inhibitor screening, singlecell tracking of dynamic FTO nuclear translocation upon starvation stimuli, FTO characterization in a biomimetic heterotypic threedimensional (3D) multicellular spheroid model, as well as the first report on the in vivo imaging of FTO activity. This strategy provides a simple yet versatile toolbox for clinical diagnosis, drug discovery, therapeutic evaluation, and biological study of RNA demethylation.

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A Demethylation-Switchable Aptamer Design Enables Lag-Free Monitoring of m⁶A Demethylase FTO with Energy Self-Sufficient and Structurally Integrated Features

Shi,

Lei,

Chen

et al. 2024

J. Am. Chem. Soc.

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show abstract

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A far red emissive RNA aptamer–fluorophore system for demethylase FTO detection: design and optimization

Abstract: Fat mass and obesity-associated protein (FTO) is a demethylase that participates in N6-methyladenosine (m6A) RNA modification. This demethylase previously linked to obesity and diabetes, is dysregulated in a number of...

Cited by 1 publication

References 44 publications

A Demethylation-Switchable Aptamer Design Enables Lag-Free Monitoring of m⁶A Demethylase FTO with Energy Self-Sufficient and Structurally Integrated Features

A Demethylation-Switchable Aptamer Design Enables Lag-Free Monitoring of m⁶A Demethylase FTO with Energy Self-Sufficient and Structurally Integrated Features

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A far red emissive RNA aptamer–fluorophore system for demethylase FTO detection: design and optimization

Abstract: Fat mass and obesity-associated protein (FTO) is a demethylase that participates in N6-methyladenosine (m6A) RNA modification. This demethylase previously linked to obesity and diabetes, is dysregulated in a number of...

Cited by 1 publication

References 44 publications

A Demethylation-Switchable Aptamer Design Enables Lag-Free Monitoring of m6A Demethylase FTO with Energy Self-Sufficient and Structurally Integrated Features

A Demethylation-Switchable Aptamer Design Enables Lag-Free Monitoring of m6A Demethylase FTO with Energy Self-Sufficient and Structurally Integrated Features

Contact Info

Product

Resources

About

A Demethylation-Switchable Aptamer Design Enables Lag-Free Monitoring of m⁶A Demethylase FTO with Energy Self-Sufficient and Structurally Integrated Features

A Demethylation-Switchable Aptamer Design Enables Lag-Free Monitoring of m⁶A Demethylase FTO with Energy Self-Sufficient and Structurally Integrated Features