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Three-dimensional DNA nanoamplifiers actuated by demethylase-activated deoxyribozyme for the ultrasensitive detection of FTO in human breast tissues.

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Title: Three-dimensional DNA nanoamplifiers actuated by demethylase-activated deoxyribozyme for the ultrasensitive detection of FTO in human breast tissues.
Authors: Fan, Li-Ping1 (AUTHOR), Wang, Xing-Miao2 (AUTHOR), Li, Juan1 (AUTHOR), Shao, Tong1 (AUTHOR), Wang, Ya-Xin1 (AUTHOR) yaxinwang@bzmc.edu.cn, Kong, De-Ming3 (AUTHOR)
Source: Sensors & Actuators B: Chemical. May2025, Vol. 431, pN.PAG-N.PAG. 1p.
Subjects: Fluorescence resonance energy transfer, Drug discovery, Gene expression, Deoxyribozymes, Adipose tissues
Abstract: Fat mass and obesity-associated protein (FTO) plays a crucial role in various physiological processes by dynamically regulating mRNA expression through the control of N6-methyladenosine (m6A) levels. Dysfunctions in FTO lead to numerous human diseases, attracting significant attention in biology and medicine. Herein, we have developed a ratiometric fluorescence biosensor based on demethylase-activated deoxyribozyme (DNAzyme) triggered three-dimensional DNA nanoamplifiers (TDNs) assisted by magnetic beads (MBs). Specifically, we engineered an m6A-silenced DNAzyme as the biosensor switch that can be specifically activated by the demethylation activity of FTO. The activated DNAzyme catalyzes the cleavage of substrates on MBs, generating multiple primers that can induce a DNA tetrahedron-mediated hyperbranched hybridization chain reaction (TD-HCR) after magnetic separation, resulting in the formation of hyperbranched products with greatly enhanced fluorescence resonance energy transfer (FRET) signals. Our approach allows for sensitive and quantitative detection of FTO with an impressive limit of detection (LOD) as low as 2.38 fM, surpassing the sensitivity of conventional ELISA kits and mostly reported methods. Furthermore, the proposed strategy enables effectively distinguishing the expression levels of FTO in tissues derived from both healthy individuals and breast cancer patients, thus offering an innovative foundation for drug discovery endeavors, investigations into m6A modifications, and advancements in clinical diagnostic capabilities. [Display omitted] • A three-dimensional DNA nanoamplifiers-based biosensor was developed for FTO analysis. • This biosensor enables triple signal amplification in an isothermal process without enzymatic participation. • This biosensor exhibits high sensitivity and good specificity. • This biosensor can work well for FTO inhibitor screening. • This biosensor can accurately quantify FTO activity in cancer cells and human breast cancer tissues. [ABSTRACT FROM AUTHOR]
Copyright of Sensors & Actuators B: Chemical is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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  Data: Three-dimensional DNA nanoamplifiers actuated by demethylase-activated deoxyribozyme for the ultrasensitive detection of FTO in human breast tissues.
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  Data: <searchLink fieldCode="AR" term="%22Fan%2C+Li-Ping%22">Fan, Li-Ping</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Wang%2C+Xing-Miao%22">Wang, Xing-Miao</searchLink><relatesTo>2</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Li%2C+Juan%22">Li, Juan</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Shao%2C+Tong%22">Shao, Tong</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Wang%2C+Ya-Xin%22">Wang, Ya-Xin</searchLink><relatesTo>1</relatesTo> (AUTHOR)<i> yaxinwang@bzmc.edu.cn</i><br /><searchLink fieldCode="AR" term="%22Kong%2C+De-Ming%22">Kong, De-Ming</searchLink><relatesTo>3</relatesTo> (AUTHOR)
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  Data: <searchLink fieldCode="JN" term="%22Sensors+%26+Actuators+B%3A+Chemical%22">Sensors & Actuators B: Chemical</searchLink>. May2025, Vol. 431, pN.PAG-N.PAG. 1p.
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  Data: <searchLink fieldCode="DE" term="%22Fluorescence+resonance+energy+transfer%22">Fluorescence resonance energy transfer</searchLink><br /><searchLink fieldCode="DE" term="%22Drug+discovery%22">Drug discovery</searchLink><br /><searchLink fieldCode="DE" term="%22Gene+expression%22">Gene expression</searchLink><br /><searchLink fieldCode="DE" term="%22Deoxyribozymes%22">Deoxyribozymes</searchLink><br /><searchLink fieldCode="DE" term="%22Adipose+tissues%22">Adipose tissues</searchLink>
– Name: Abstract
  Label: Abstract
  Group: Ab
  Data: Fat mass and obesity-associated protein (FTO) plays a crucial role in various physiological processes by dynamically regulating mRNA expression through the control of N6-methyladenosine (m6A) levels. Dysfunctions in FTO lead to numerous human diseases, attracting significant attention in biology and medicine. Herein, we have developed a ratiometric fluorescence biosensor based on demethylase-activated deoxyribozyme (DNAzyme) triggered three-dimensional DNA nanoamplifiers (TDNs) assisted by magnetic beads (MBs). Specifically, we engineered an m6A-silenced DNAzyme as the biosensor switch that can be specifically activated by the demethylation activity of FTO. The activated DNAzyme catalyzes the cleavage of substrates on MBs, generating multiple primers that can induce a DNA tetrahedron-mediated hyperbranched hybridization chain reaction (TD-HCR) after magnetic separation, resulting in the formation of hyperbranched products with greatly enhanced fluorescence resonance energy transfer (FRET) signals. Our approach allows for sensitive and quantitative detection of FTO with an impressive limit of detection (LOD) as low as 2.38 fM, surpassing the sensitivity of conventional ELISA kits and mostly reported methods. Furthermore, the proposed strategy enables effectively distinguishing the expression levels of FTO in tissues derived from both healthy individuals and breast cancer patients, thus offering an innovative foundation for drug discovery endeavors, investigations into m6A modifications, and advancements in clinical diagnostic capabilities. [Display omitted] • A three-dimensional DNA nanoamplifiers-based biosensor was developed for FTO analysis. • This biosensor enables triple signal amplification in an isothermal process without enzymatic participation. • This biosensor exhibits high sensitivity and good specificity. • This biosensor can work well for FTO inhibitor screening. • This biosensor can accurately quantify FTO activity in cancer cells and human breast cancer tissues. [ABSTRACT FROM AUTHOR]
– Name: AbstractSuppliedCopyright
  Label:
  Group: Ab
  Data: <i>Copyright of Sensors & Actuators B: Chemical is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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        Value: 10.1016/j.snb.2025.137431
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        Text: English
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      – SubjectFull: Drug discovery
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      – SubjectFull: Gene expression
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      – SubjectFull: Deoxyribozymes
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      – SubjectFull: Adipose tissues
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      – TitleFull: Three-dimensional DNA nanoamplifiers actuated by demethylase-activated deoxyribozyme for the ultrasensitive detection of FTO in human breast tissues.
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            NameFull: Fan, Li-Ping
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            NameFull: Wang, Xing-Miao
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            NameFull: Wang, Ya-Xin
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              M: 05
              Text: May2025
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              Y: 2025
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