Wei Yu, Xu Xuehui, Shang Yingxu, Jiang Qiao, Li Can, Ding Baoquan
Bio-X Institutes, Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders (Ministry of Education), Shanghai Jiao Tong University 200030 Shanghai China.
CAS Key Laboratory of Nanosystem and Hierarchical Fabrication, CAS Center for Excellence in Nanoscience, National Center for NanoScience and Technology 11 BeiYiTiao, ZhongGuanCun 100190 Beijing China
RSC Adv. 2019 May 15;9(27):15205-15209. doi: 10.1039/c9ra01769k. eCollection 2019 May 14.
Rolling circle amplification (RCA) and rolling circle transcription (RCT) can be used to fabricate various structures and organize functional materials for biological applications. The full understanding of the interactions between RCA/RCT-derived structures and live cells is urgently demanded. Here, we present a label-free fluorescent strategy to study the intracellular location and stability of RCA-based DNA flowers in live cells. The DNA flower structures are co-assembled with carbazole-based biscyanine fluorophores, which are DNA detecting molecules and characterized by restriction of intramolecular rotation (RIR) induced strong fluorescent emission. When biscyanine molecules are encapsulated in the DNA flowers electrostatic attraction, these confined RIR dyes can produce strong luminescent emission. Using this advantage, we use the RIR enhanced technique for direct visualization of the distribution and degradation of DNA flowers in live cellular systems. Our current research could be adapted to other advanced DNA-based materials, providing a new strategy to fabricate fluorescent DNA materials and realize controllable release of payloads.
滚环扩增(RCA)和滚环转录(RCT)可用于构建各种结构并组织用于生物应用的功能材料。目前迫切需要全面了解RCA/RCT衍生结构与活细胞之间的相互作用。在此,我们提出了一种无标记荧光策略,用于研究活细胞中基于RCA的DNA花的细胞内定位和稳定性。DNA花结构与基于咔唑的双氰基荧光团共同组装,这些荧光团是DNA检测分子,其特征在于分子内旋转受限(RIR)诱导强烈的荧光发射。当双氰基分子通过静电吸引被包裹在DNA花中时,这些受限的RIR染料可以产生强烈的发光发射。利用这一优势,我们使用RIR增强技术直接可视化活细胞系统中DNA花的分布和降解。我们目前的研究可以应用于其他先进的基于DNA的材料,为制造荧光DNA材料和实现有效载荷的可控释放提供了一种新策略。