Shah Pratik, Thulstrup Peter Waaben, Cho Seok Keun, Bhang Yong-Joo, Ahn Jong Cheol, Choi Suk Won, Bjerrum Morten Jannik, Yang Seong Wook
UNIK Center for Synthetic Biology/Plant Biochemistry Laboratory, University of Copenhagen, Thorvaldsensvej 40, DK-1871 Frederiksberg C, Copenhagen, Denmark.
Analyst. 2014 May 7;139(9):2158-66. doi: 10.1039/c3an02150e.
MicroRNAs (miRNAs) are small regulatory RNAs (size ∼21nt to ∼25nt) that can be used as biomarkers of disease diagnosis, and efforts have been directed towards the invention of a rapid, simple and sequence-selective detection method for miRNAs. We recently developed a DNA/silver nanoclusters (AgNCs)-based turn-off fluorescence method in the presence of target miRNA. To further advance our method toward multiplex miRNA detection in solution, the design of various fluorescent DNA/AgNCs probes was essential. Therefore, tethering of DNA-12nt scaffolds with 9 different AgNCs emitters to target-sensing DNA sequences was investigated. Interestingly, for the creation of spectrally different DNA/AgNCs probes, not only were the emitters encapsulated in 9 different DNA-12nt scaffolds necessary but the tethered target-sensing DNA sequences are also crucial to tune the fluorescence across the visible to infra-red region. In this study, we obtained three spectrally distinctive emitters of each DNA/AgNCs probes such as green, red, and near-infrared (NIR) fluorescence. Using these DNA/AgNCs probes, we here show a proof of concept for a rapid, one-step, in-solution multiplex miRNA detection method.
微小RNA（miRNA）是一类小的调节性RNA（大小约为21nt至25nt），可作为疾病诊断的生物标志物，人们一直致力于发明一种快速、简单且具有序列选择性的miRNA检测方法。我们最近开发了一种基于DNA/银纳米簇（AgNCs）的猝灭荧光方法，用于检测目标miRNA。为了进一步将我们的方法推进到溶液中的多重miRNA检测，设计各种荧光DNA/AgNCs探针至关重要。因此，研究了将具有9种不同AgNCs发射体的DNA-12nt支架与目标传感DNA序列相连。有趣的是，为了创建光谱不同的DNA/AgNCs探针，不仅将发射体封装在9种不同的DNA-12nt支架中是必要的，而且相连的目标传感DNA序列对于调节从可见光到红外区域的荧光也至关重要。在本研究中，我们获得了每种DNA/AgNCs探针的三种光谱独特的发射体，如绿色、红色和近红外（NIR）荧光。使用这些DNA/AgNCs探针，我们在此展示了一种快速、一步、溶液中多重miRNA检测方法的概念验证。
