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

被引：0

作者：

Shi, Yakun ^{[1
]}

Lei, Yutian ^{[1
]}

Chen, Meng ^{[1
]}

Ma, Hansu ^{[2
]}

Shen, Taorong ^{[1
]}

Zhang, Yanfei ^{[1
]}

Huang, Xing ^{[1
]}

Ling, Wanxuan ^{[1
]}

Liu, Si-Yang ^{[1
]}

Pan, Yihang ^{[2
]}

Dai, Zong ^{[1
]}

Xu, Yuzhi ^{[2
]}

机构：

[1] Sun Yat Sen Univ, Sch Biomed Engn, Guangdong Prov Key Lab Sensing Technol & Biomed In, Shenzhen Campus, Shenzhen 518107, Peoples R China

[2] Sun Yat Sen Univ, Affiliated Hosp 7, Precis Med Ctr, Guangdong Prov Key Lab Digest Canc Res, Shenzhen 518107, Peoples R China

来源：

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY | 2024年 / 146卷 / 50期

基金：

中国国家自然科学基金;

关键词：

NUCLEAR TRANSLOCATION; RNA; SUBSTRATE;

D O I：

10.1021/jacs.4c12884

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Cellular context profiling of modification effector proteins is critical for an in-depth understanding of their biological roles in RNA N 6-methyladenosine (m6A) 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 demethylation-switchable aptamer sensor engineered with a site-specific m6A (DSA-m6A) for lag-free monitoring of the m6A demethylase FTO activity in living cells. As a proof of concept, a DNA aptamer against adenosine triphosphate (ATP) is selected to construct the DSA-m6A model, as the "universal energy currency" role of ATP could guarantee the equally fast and spontaneous conformation change of DSA-m6A 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-m6A design facilitates biomedical research, including live-cell imaging, inhibitor screening, single-cell tracking of dynamic FTO nuclear translocation upon starvation stimuli, FTO characterization in a biomimetic heterotypic three-dimensional (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.

引用

页码：34638 / 34650

页数：13