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Ratiometric Fluorescence Sensor for the MicroRNA Determination by Catalyzed Hairpin Assembly
ACS Sensors ( IF 8.2 ) Pub Date : 2017-10-09 00:00:00 , DOI: 10.1021/acssensors.7b00313 Yi Liu 1 , Tian Shen 1 , Jing Li 1 , Hang Gong 1 , Chunyan Chen 1 , Xiaoming Chen 1 , Changqun Cai 1
ACS Sensors ( IF 8.2 ) Pub Date : 2017-10-09 00:00:00 , DOI: 10.1021/acssensors.7b00313 Yi Liu 1 , Tian Shen 1 , Jing Li 1 , Hang Gong 1 , Chunyan Chen 1 , Xiaoming Chen 1 , Changqun Cai 1
Affiliation
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A novel catalyzed hairpin assembly-based turn-on ratiometric fluorescence biosensor was constructed for the determination of microRNA-122 (miRNA-122) by using 2-aminopurine (2-AP) and thioflavin T (ThT) as detection signal sources. Hairpin DNA sequence (H1) includes the complementary strands of miRNA-122 and G-quadruplex-forming sequence. When miRNA-122 was presented, hybridization occurred between miRNA-122 and part of H1, causing a double-stranded DNA and a G-quadruplex formed. The formed double-stranded DNA significantly decreased the fluorescence intensity of 2-AP. Furthermore, after binding with ThT, the formed G-quadruplex led to the fluorescent enhancement. The hairpin DNA sequence (H2) hybridized with the unfolded H1 and displaced miRNA-122. Finally, the displaced miRNA-122 again hybridized with the H1 and initiated cycle amplification. This sensor showed a linear ranges of 0.5–50 nM and the limit of detection for miRNA-122 assay was 72 pM (with the lowest measured concentration of 500 pM) for determination of miRNA-122 when no other miRNA was present. Measurements on cell lysates from 100, 1000, and 10 000 cells of three different cell lines provided increasing signal ratios, which showed the application potential of the sensor for miRNA determination in real samples.
中文翻译：
用于催化发夹装配测定MicroRNA的比例荧光传感器
通过使用2-氨基嘌呤（2-AP）和硫代黄素T（ThT）作为检测信号源，构建了一种基于催化发夹装配的新型开启比例荧光生物传感器，用于测定microRNA-122（miRNA-122）。发夹DNA序列（H1）包括miRNA-122和G-四链体形成序列的互补链。提出miRNA-122时，miRNA-122与H1的一部分发生杂交，导致双链DNA形成G-四链体。形成的双链DNA显着降低了2-AP的荧光强度。此外，在与ThT结合后，形成的G-四链体导致荧光增强。发夹DNA序列（H2）与未折叠的H1和置换的miRNA-122杂交。最终，置换的miRNA-122再次与H1杂交并开始循环扩增。该传感器显示0.5–50 nM的线性范围，当不存在其他miRNA时，用于miRNA-122测定的miRNA-122检测限为72 pM（最低测量浓度为500 pM）。对三种不同细胞系的100、1000和10000个细胞的细胞裂解液进行的测量提供了增加的信号比率，这表明传感器在实际样品中用于miRNA测定的应用潜力。
更新日期：2017-10-09
中文翻译：
用于催化发夹装配测定MicroRNA的比例荧光传感器
通过使用2-氨基嘌呤（2-AP）和硫代黄素T（ThT）作为检测信号源，构建了一种基于催化发夹装配的新型开启比例荧光生物传感器，用于测定microRNA-122（miRNA-122）。发夹DNA序列（H1）包括miRNA-122和G-四链体形成序列的互补链。提出miRNA-122时，miRNA-122与H1的一部分发生杂交，导致双链DNA形成G-四链体。形成的双链DNA显着降低了2-AP的荧光强度。此外，在与ThT结合后，形成的G-四链体导致荧光增强。发夹DNA序列（H2）与未折叠的H1和置换的miRNA-122杂交。最终，置换的miRNA-122再次与H1杂交并开始循环扩增。该传感器显示0.5–50 nM的线性范围，当不存在其他miRNA时，用于miRNA-122测定的miRNA-122检测限为72 pM（最低测量浓度为500 pM）。对三种不同细胞系的100、1000和10000个细胞的细胞裂解液进行的测量提供了增加的信号比率，这表明传感器在实际样品中用于miRNA测定的应用潜力。
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