Roknabadi Nastaran, Borghei Yasaman-Sadat, Seifezadeh Seyedeh Saina, Soltani Bahram M, Mowla Seyed Javad
Department of Molecular Genetics Faculty of Biological Sciences, Tarbiat Modares University, Tehran 14115-175, Iran.
Center for Bioscience & Technology, Institute for Convergence Science & Technology Sharif University of Technology, Tehran 1458889694, Iran.
ACS Omega. 2024 Aug 21;9(35):37205-37212. doi: 10.1021/acsomega.4c04565. eCollection 2024 Sep 3.
The application of nanobiotechnology in biomolecule detection can provide fast and accurate tests for diagnosing molecular changing-associated diseases. The use of AuNPs-thiolated probe conjugates has long been considered as an alternative method for the detection of specific DNA/RNA targets. Here, we present a colorimetric direct detection method for the SOX2OT transcript, long noncoding RNAs (lncRNAs), by using a poly guanine tail (G12) as a template for in situ synthesis of gold nanoparticles (AuNPs) without any chemical modification or DNA labeling. We have then developed this proposed detection system based on two complementary sequences of long noncoding RNA SOX2OT with an extra strand of poly G12. Using this method, we were able to differentiate lung squamous cell carcinoma from adenocarcinoma samples. Based on this disclosure, this invention provides a simple visual method to detect specific lncRNA sequences without the need for amplifying the target lncRNA and discriminate squamous cell carcinoma from adenocarcinoma samples. Our invention provides a diagnostic kit to detect RNA by means of direct detection (PCR-free) of the lncRNA by in situ synthesis of AuNPs based on two probes with an extra strand of poly G12.
纳米生物技术在生物分子检测中的应用可为诊断与分子变化相关的疾病提供快速准确的检测方法。长期以来，金纳米颗粒-硫醇化探针缀合物的使用一直被视为检测特定DNA/RNA靶标的替代方法。在此，我们提出了一种用于检测SOX2OT转录本（一种长链非编码RNA，即lncRNA）的比色直接检测方法，该方法使用聚鸟嘌呤尾（G12）作为原位合成金纳米颗粒（AuNPs）的模板，无需任何化学修饰或DNA标记。然后，我们基于长链非编码RNA SOX2OT的两个互补序列以及一条额外的聚G12链开发了该检测系统。使用这种方法，我们能够区分肺鳞状细胞癌和腺癌样本。基于本公开内容，本发明提供了一种简单的可视化方法，无需扩增目标lncRNA即可检测特定的lncRNA序列，并区分鳞状细胞癌和腺癌样本。本发明提供了一种诊断试剂盒，通过基于带有额外聚G12链的两个探针原位合成AuNPs来直接检测（无需PCR）lncRNA从而检测RNA。
