Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Classical Triplex Molecular Beacons for MicroRNA-21 and Vascular Endothelial Growth Factor Detection
ACS Sensors ( IF 8.2 ) Pub Date : 2018-10-17 00:00:00 , DOI: 10.1021/acssensors.8b00996 Shasha Lu 1, 2 , Shuang Wang 1, 2 , Jiahui Zhao 1, 3 , Jian Sun 1 , Xiurong Yang 1, 2
ACS Sensors ( IF 8.2 ) Pub Date : 2018-10-17 00:00:00 , DOI: 10.1021/acssensors.8b00996 Shasha Lu 1, 2 , Shuang Wang 1, 2 , Jiahui Zhao 1, 3 , Jian Sun 1 , Xiurong Yang 1, 2
Affiliation
|
Triplex molecular beacons (tMBs) possess great potential in biological sensing because of the pH responsiveness and controllability of binding strength. Here, we systematically investigate and rationally design a classical tMB for convenient detection of microRNA-21, a well-known biomarker of cardio-cerebrovascular diseases. In the tMB, we employ the complementary sequence of miR-21 as the loop and the sequences of protonated cytosine–guanine–cytosine (C-G•C+) and thymine–adenine–thymine (T-A•T) as the triplex stem, in which both the Watson–Crick and Hoogsteen base-pairing control the binding strength in cooperation. It is demonstrated for the first time that the presence of miR-21 would only break the Hoogsteen base-pairing in the stem and hybridize with the tMB to form the rigid heterozygous hybrid duplex structure. These would hinder the fluorescence resonance energy transfer (FRET) between the fluorophore (FAM) and quencher (BHQ1) labeled at the ends of the oligonucleotide, and the fluorescence recovery degree of FAM can be used as the standard to quantitate the miR-21. More significantly, the excellent adjustability and sensitivity of our tMBs have been confirmed by constructing the corresponding duplex molecular beacon (dMB) for comparison. The fluorophore FAM in the tMB could be replaced by the fluorescent DNA/silver nanoclusters, which exhibits the universal applicability of energy donor and receptor selection for tMB. Furthermore, our proposed tMB could also be developed as an aptasensor for the detection of vascular endothelial growth factor (VEGF) by only introducing the complementary sequence of its aptamer into the tMB. This work is of great significance for the systematic study of tMBs for the detection of biomarkers such as nucleic acids and proteins.
中文翻译：
用于MicroRNA-21和血管内皮生长因子检测的经典三链体分子信标
三重分子信标（tMBs）由于pH响应能力和结合强度的可控性，在生物传感方面具有巨大潜力。在这里，我们系统地研究和合理设计了经典的tMB，以方便检测microRNA-21（一种心脑血管疾病的著名生物标志物）。在tMB中，我们将miR-21的互补序列用作环，将质子化的胞嘧啶-鸟嘌呤-胞嘧啶（CG•C +）和胸腺嘧啶-腺嘌呤-胸腺嘧啶（TA•T）序列用作三链体茎，其中两者Watson-Crick和Hoogsteen碱基配对控制着结合力。首次证明，miR-21的存在只会破坏茎中的Hoogsteen碱基配对，并与tMB杂交形成刚性的杂合杂合双链体结构。这些会阻碍在寡核苷酸末端标记的荧光团（FAM）和淬灭剂（BHQ1）之间的荧光共振能量转移（FRET），并且FAM的荧光恢复度可以用作定量miR-21的标准。更重要的是，通过构建相应的双链分子信标（dMB）进行比较，我们的tMB具有出色的可调节性和敏感性。tMB中的荧光团FAM可以用荧光DNA /银纳米团簇代替，这显示了tMB能量供体和受体选择的普遍适用性。此外，我们提出的tMB还可以通过仅将适体的互补序列引入tMB中而开发为用于检测血管内皮生长因子（VEGF）的适体传感器。
更新日期：2018-10-17
中文翻译：
用于MicroRNA-21和血管内皮生长因子检测的经典三链体分子信标
三重分子信标（tMBs）由于pH响应能力和结合强度的可控性，在生物传感方面具有巨大潜力。在这里，我们系统地研究和合理设计了经典的tMB，以方便检测microRNA-21（一种心脑血管疾病的著名生物标志物）。在tMB中，我们将miR-21的互补序列用作环，将质子化的胞嘧啶-鸟嘌呤-胞嘧啶（CG•C +）和胸腺嘧啶-腺嘌呤-胸腺嘧啶（TA•T）序列用作三链体茎，其中两者Watson-Crick和Hoogsteen碱基配对控制着结合力。首次证明，miR-21的存在只会破坏茎中的Hoogsteen碱基配对，并与tMB杂交形成刚性的杂合杂合双链体结构。这些会阻碍在寡核苷酸末端标记的荧光团（FAM）和淬灭剂（BHQ1）之间的荧光共振能量转移（FRET），并且FAM的荧光恢复度可以用作定量miR-21的标准。更重要的是，通过构建相应的双链分子信标（dMB）进行比较，我们的tMB具有出色的可调节性和敏感性。tMB中的荧光团FAM可以用荧光DNA /银纳米团簇代替，这显示了tMB能量供体和受体选择的普遍适用性。此外，我们提出的tMB还可以通过仅将适体的互补序列引入tMB中而开发为用于检测血管内皮生长因子（VEGF）的适体传感器。
京公网安备 11010802027423号