OWL2: a molecular beacon-based nanostructure for highly selective detection of single-nucleotide variations in folded nucleic acids
OWL2: a molecular beacon-based nanostructure for highly selective detection of single-nucleotide variations in folded nucleic acids
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DOI：
10.1039/d2nr05590b
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发表时间：
2023-02-27
期刊：
影响因子：
6.7
通讯作者：
Kolpashchikov，Dmitry M.
中科院分区：
文献类型：
--
作者：
Mueller，Brittany L.;Liberman，Mark J.;Kolpashchikov，Dmitry M.
在过去的50年中，杂交探针已被用于检测特异性核酸。尽管进行了大量的努力并且意义重大，但常用探针的挑战包括（1）在低温（例如室温或37 °C）下检测单核苷酸变异（SNV）的低选择性;（2）结合折叠核酸的低亲和力;以及（3）荧光探针的成本。在这里，我们介绍了一种多组分杂交探针，称为OWL 2传感器，它解决了所有这三个问题。OWL 2传感器使用两个分析物结合臂来紧密结合和解开折叠的分析物，以及两个结合分析物和通用分子信标（UMB）探针以形成荧光“OWL”结构的序列特异性链。OWL 2传感器能够在5-38 °C的温度范围内区分折叠分析物中的单碱基错配。该设计具有成本效益，因为相同的UMB探针可用于检测任何分析物序列。
Hybridization probes have been used in the detection of specific nucleic acids for the last 50 years. Despite the extensive efforts and the great significance, the challenges of the commonly used probes include (1) low selectivity in detecting single nucleotide variations (SNV) at low (e.g. room or 37 °C) temperatures; (2) low affinity in binding folded nucleic acids, and (3) the cost of fluorescent probes. Here we introduce a multicomponent hybridization probe, called OWL2 sensor, which addresses all three issues. The OWL2 sensor uses two analyte binding arms to tightly bind and unwind folded analytes, and two sequence-specific strands that bind both the analyte and a universal molecular beacon (UMB) probe to form fluorescent ‘OWL’ structure. The OWL2 sensor was able to differentiate single base mismatches in folded analytes in the temperature range of 5–38 °C. The design is cost-efficient since the same UMB probe can be used for detecting any analyte sequence.