Dual Catalytic Hairpin Assembly-Based Automatic Molecule Machine for Amplified Detection of Auxin Response Factor-Targeted MicroRNA-160

MicroRNA160 plays a crucial role in plant development by negatively regulating the auxin response factors (ARFs). In this manuscript, we design an automatic molecule machine (AMM) based on the dual catalytic hairpin assembly (D-CHA) strategy for the signal amplification detection of miRNA160. The de...

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Autores principales:	Lei Wang, Xing Dai, Yujian Feng, Qiyang Zhao, Lin Liu, Chang Xue, Langtao Xiao, Ruozhong Wang
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	miRNA160 dual catalytic hairpin assembly process signal amplification strategy AMM system strand displacement reaction Organic chemistry QD241-441
Acceso en línea:	https://doaj.org/article/4100868e32394a2e9deb910585150aa5
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spelling	oai:doaj.org-article:4100868e32394a2e9deb910585150aa52021-11-11T18:26:54ZDual Catalytic Hairpin Assembly-Based Automatic Molecule Machine for Amplified Detection of Auxin Response Factor-Targeted MicroRNA-16010.3390/molecules262164321420-3049https://doaj.org/article/4100868e32394a2e9deb910585150aa52021-10-01T00:00:00Zhttps://www.mdpi.com/1420-3049/26/21/6432https://doaj.org/toc/1420-3049MicroRNA160 plays a crucial role in plant development by negatively regulating the auxin response factors (ARFs). In this manuscript, we design an automatic molecule machine (AMM) based on the dual catalytic hairpin assembly (D-CHA) strategy for the signal amplification detection of miRNA160. The detection system contains four hairpin-shaped DNA probes (HP1, HP2, HP3, and HP4). For HP1, the loop is designed to be complementary to miRNA160. A fragment of DNA with the same sequences as miRNA160 is separated into two pieces that are connected at the 3′ end of HP2 and 5′ end of HP3, respectively. In the presence of the target, four HPs are successively dissolved by the first catalytic hairpin assembly (CHA1), forming a four-way DNA junction (F-DJ) that enables the rearrangement of separated DNA fragments at the end of HP2 and HP3 and serving as an integrated target analogue for initiating the second CHA reaction, generating an enhanced fluorescence signal. Assay experiments demonstrate that D-CHA has a better performance compared with traditional CHA, achieving the detection limit as low as 10 pM for miRNA160 as deduced from its corresponding DNA surrogates. Moreover, non-target miRNAs, as well as single-base mutation targets, can be detected. Overall, the D-CHA strategy provides a competitive method for plant miRNAs detection.Lei WangXing DaiYujian FengQiyang ZhaoLin LiuChang XueLangtao XiaoRuozhong WangMDPI AGarticlemiRNA160dual catalytic hairpin assembly processsignal amplification strategyAMM systemstrand displacement reactionOrganic chemistryQD241-441ENMolecules, Vol 26, Iss 6432, p 6432 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	miRNA160 dual catalytic hairpin assembly process signal amplification strategy AMM system strand displacement reaction Organic chemistry QD241-441
spellingShingle	miRNA160 dual catalytic hairpin assembly process signal amplification strategy AMM system strand displacement reaction Organic chemistry QD241-441 Lei Wang Xing Dai Yujian Feng Qiyang Zhao Lin Liu Chang Xue Langtao Xiao Ruozhong Wang Dual Catalytic Hairpin Assembly-Based Automatic Molecule Machine for Amplified Detection of Auxin Response Factor-Targeted MicroRNA-160
description	MicroRNA160 plays a crucial role in plant development by negatively regulating the auxin response factors (ARFs). In this manuscript, we design an automatic molecule machine (AMM) based on the dual catalytic hairpin assembly (D-CHA) strategy for the signal amplification detection of miRNA160. The detection system contains four hairpin-shaped DNA probes (HP1, HP2, HP3, and HP4). For HP1, the loop is designed to be complementary to miRNA160. A fragment of DNA with the same sequences as miRNA160 is separated into two pieces that are connected at the 3′ end of HP2 and 5′ end of HP3, respectively. In the presence of the target, four HPs are successively dissolved by the first catalytic hairpin assembly (CHA1), forming a four-way DNA junction (F-DJ) that enables the rearrangement of separated DNA fragments at the end of HP2 and HP3 and serving as an integrated target analogue for initiating the second CHA reaction, generating an enhanced fluorescence signal. Assay experiments demonstrate that D-CHA has a better performance compared with traditional CHA, achieving the detection limit as low as 10 pM for miRNA160 as deduced from its corresponding DNA surrogates. Moreover, non-target miRNAs, as well as single-base mutation targets, can be detected. Overall, the D-CHA strategy provides a competitive method for plant miRNAs detection.
format	article
author	Lei Wang Xing Dai Yujian Feng Qiyang Zhao Lin Liu Chang Xue Langtao Xiao Ruozhong Wang
author_facet	Lei Wang Xing Dai Yujian Feng Qiyang Zhao Lin Liu Chang Xue Langtao Xiao Ruozhong Wang
author_sort	Lei Wang
title	Dual Catalytic Hairpin Assembly-Based Automatic Molecule Machine for Amplified Detection of Auxin Response Factor-Targeted MicroRNA-160
title_short	Dual Catalytic Hairpin Assembly-Based Automatic Molecule Machine for Amplified Detection of Auxin Response Factor-Targeted MicroRNA-160
title_full	Dual Catalytic Hairpin Assembly-Based Automatic Molecule Machine for Amplified Detection of Auxin Response Factor-Targeted MicroRNA-160
title_fullStr	Dual Catalytic Hairpin Assembly-Based Automatic Molecule Machine for Amplified Detection of Auxin Response Factor-Targeted MicroRNA-160
title_full_unstemmed	Dual Catalytic Hairpin Assembly-Based Automatic Molecule Machine for Amplified Detection of Auxin Response Factor-Targeted MicroRNA-160
title_sort	dual catalytic hairpin assembly-based automatic molecule machine for amplified detection of auxin response factor-targeted microrna-160
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/4100868e32394a2e9deb910585150aa5
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_version_	1718431818815897600

Dual Catalytic Hairpin Assembly-Based Automatic Molecule Machine for Amplified Detection of Auxin Response Factor-Targeted MicroRNA-160

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