Department of Molecular Biology, ICMR-National Institute for Research in Environmental Health, Bhopal, India.
Department of General and Inorganic Chemistry, Saratov State University, Saratov, Russia.
Nanomedicine. 2021 Aug;36:102413. doi: 10.1016/j.nano.2021.102413. Epub 2021 Jun 17.
Development of a rapid, sensitive and easy to use point of care assay for detection of circulating long non-coding RNAs (lncRNAs) is of great importance. These biomolecules possess the ability to regulate vital cellular processes and act as biomarkers for various human non-communicable diseases. The present work aimed to develop a simplified and reliable cytometric fluorescence-based approach for precise recognition of circulating lncRNAs in a given sample using biotinylated uracil-modified oligonucleotide tethered AlexaFluor488-labeled streptavidin gold colloidal (BiO-StrAG) nano-conjugates. The fluorophores in close proximity to the gold nanoparticles result in quenching of fluorescence; however, specific recognition of target lncRNAs increases this distance which causes plasmonic enhancement of fluorescence. As per the flow cytometry and fluorometry investigations, the developed methodology provides a precise and sensitive approach for detection of the target lncRNAs (up to 5 nM in any given sample). With advantages of high selectivity and feasibility, our strategy offers great potential of being developed as a promising tool for interrogating aberrant regulation of lncRNAs functions, especially indicated in various diseased states.
开发一种快速、灵敏且易于使用的即时检测分析方法来检测循环长非编码 RNA（lncRNA）非常重要。这些生物分子具有调节重要细胞过程的能力，并可作为各种人类非传染性疾病的生物标志物。本工作旨在开发一种简化且可靠的基于细胞仪荧光的方法，使用生物素化尿嘧啶修饰的寡核苷酸连接的 AlexaFluor488 标记链霉亲和素金纳米胶体（BiO-StrAG）纳米缀合物，精确识别给定样品中的循环 lncRNA。靠近金纳米粒子的荧光团会导致荧光猝灭；然而，目标 lncRNA 的特异性识别会增加这种距离，从而导致荧光的等离子体增强。根据流式细胞术和荧光计的研究，所开发的方法学提供了一种精确和灵敏的方法来检测靶 lncRNA（在任何给定的样品中可达 5 nM）。该策略具有高选择性和可行性的优势，有望成为研究 lncRNA 功能异常调节的有前途的工具，特别是在各种疾病状态下。
