Afonin Kirill A, Schultz Danielle, Jaeger Luc, Gwinn Elisabeth, Shapiro Bruce A
Center for Cancer Research Nanobiology Program, National Cancer Institute, Frederick, MD, 21702, USA.
Methods Mol Biol. 2015;1297:59-66. doi: 10.1007/978-1-4939-2562-9_4.
The growing interest in designing functionalized, RNA-based nanoparticles (NPs) for applications such as cancer therapeutics requires simple, efficient assembly assays. Common methods for tracking RNA assemblies such as native polyacrylamide gels and atomic force microscopy are often time-intensive and, therefore, undesirable. Here we describe a technique for rapid analysis of RNA NP assembly stages using the formation of fluorescent silver nanoclusters (Ag NCs). This method exploits the single-stranded specificity and sequence dependence of Ag NC formation to produce unique optical readouts for each stage of RNA NP assembly, obtained readily after synthesis.
对于设计用于癌症治疗等应用的功能化RNA纳米颗粒(NP)的兴趣日益增长,这需要简单、高效的组装检测方法。用于追踪RNA组装体的常用方法,如天然聚丙烯酰胺凝胶和原子力显微镜,通常耗时较长,因此并不理想。在此,我们描述了一种利用荧光银纳米簇(Ag NCs)的形成来快速分析RNA NP组装阶段的技术。该方法利用了Ag NC形成的单链特异性和序列依赖性,为RNA NP组装的每个阶段产生独特的光学读数,合成后即可轻松获得。