Department of Chemical and Biomolecular Engineering, National University of Singapore, 4 Engineering Drive 4, 117585 Singapore, Singapore.
ACS Nano. 2015 May 26;9(5):5609-17. doi: 10.1021/acsnano.5b01954. Epub 2015 Apr 28.
Rapid and precise in situ detection of gene expressions within a single cell is highly informative and offers valuable insights into its state. Detecting mRNA within single cells in real time and nondestructively remains an important challenge. Using DNA nanotechnology and inspired by nature's many examples of "protective-yet-accessible" exoskeletons, we designed our mRNA nanosensor, nano-snail-inspired nucleic acid locator (nano-SNEL), to illustrate these elements. The design of the nano-SNEL is composed of a sensory molecular beacon module to detect mRNA and a DNA nanoshell component, mimicking the functional anatomy of a snail. Accurate and sensitive visualization of mRNA is achieved by the exceptional protection conferred by the nanoshell to the sensory component from nucleases-mediated degradation by approximately 9-25-fold compared to its unprotected counterpart. Our nano-SNEL design strategy improved cell internalization is a demonstration of accurate, dynamic spatiotemporal resolved detection of RNA transcripts in living cells.
快速准确地检测单个细胞内的基因表达具有重要意义，可为了解细胞状态提供有价值的信息。实时、非破坏性地检测单细胞内的 mRNA 仍然是一个重要的挑战。我们受自然界中许多“保护但可及”外骨骼的启发，利用 DNA 纳米技术设计了我们的 mRNA 纳米传感器，即纳米蜗牛启发型核酸定位器（nano-SNEL），以展示这些元素。nano-SNEL 的设计由一个感应分子信标模块组成，用于检测 mRNA，以及一个 DNA 纳米壳组件，模拟蜗牛的功能解剖结构。纳米壳为感应元件提供了极好的保护，使其免受核酸酶介导的降解，与未受保护的分子信标相比，降解效率降低了约 9-25 倍，从而实现了 mRNA 的精确和灵敏可视化。我们的 nano-SNEL 设计策略提高了细胞内化效率，展示了在活细胞中对 RNA 转录物进行准确、动态、时空分辨检测的能力。
