Using a deoxyribozyme ligase and rolling circle amplification to detect a non-nucleic acid analyte, ATP

Eun Jeonq Cho; Litao Yanq; Matthew Lew; Andrew D. Ellinqton

doi:10.1021/ja043490u

Using a deoxyribozyme ligase and rolling circle amplification to detect a non-nucleic acid analyte, ATP

Eun Jeonq Cho, Litao Yanq, Matthew Lew, Andrew D. Ellinqton

Research output: Contribution to journal › Article › peer-review

182 Scopus citations

Abstract

An allosteric ribozyme (aptazyme) has been used to transduce the binding of a small organic analyte (ATP) into the ligation of a circular template for rolling circle amplification (RCA). An ATP-activated deoxyribozyme ligase was immobilized on a glass slide and, upon addition of ATP, catalyzed the ligation of a circular padlock probe. The ligated products could be directly amplified and visualized via RCA. The coupled reaction exhibited could detect as little as 1 μM of ATP and could discriminate against structurally similar nucleotides such as GTP, CTP, and UTP. Cooperative ATP activation of the deoxyribozyme was faithfully mimicked by RCA, yielding an amplified "switch" that was responsive to ATP concentration.

Original language	English (US)
Pages (from-to)	2022-2023
Number of pages	2
Journal	Journal of the American Chemical Society
Volume	127
Issue number	7
DOIs	https://doi.org/10.1021/ja043490u
State	Published - Feb 23 2005
Externally published	Yes

ASJC Scopus subject areas

Catalysis
General Chemistry
Biochemistry
Colloid and Surface Chemistry

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AU - Lew, Matthew

AU - Ellinqton, Andrew D.

PY - 2005/2/23

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AB - An allosteric ribozyme (aptazyme) has been used to transduce the binding of a small organic analyte (ATP) into the ligation of a circular template for rolling circle amplification (RCA). An ATP-activated deoxyribozyme ligase was immobilized on a glass slide and, upon addition of ATP, catalyzed the ligation of a circular padlock probe. The ligated products could be directly amplified and visualized via RCA. The coupled reaction exhibited could detect as little as 1 μM of ATP and could discriminate against structurally similar nucleotides such as GTP, CTP, and UTP. Cooperative ATP activation of the deoxyribozyme was faithfully mimicked by RCA, yielding an amplified "switch" that was responsive to ATP concentration.

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Using a deoxyribozyme ligase and rolling circle amplification to detect a non-nucleic acid analyte, ATP

Abstract

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