We have carefully designed a probe for in situ detection and imaging of Argonaute2 (Ago2), a key RNA interference (RNAi) protein in human cells and siRNA (Let-7a) interactions. The probe mainly contains two parts: a gold nanoparticle (AuNP) being quenched and a functionalized nucleic acid as a fluorescent output carrier. After the probe is endocytosed into the cell, the modified hairpin nucleic acid can be cut into two parts by the Ago2/Let-7a complex (ALC). The cut nucleic acid will replace a DNA strand modified on another duplex nucleic acid. The replaced DNA strand’s 3’-end is modified with a fluorophore (Cy3). By cascading single cells, the probe can directly achieve the fluorescence recovery of the probe in the cell. By dynamically monitoring the in-situ quantitative response of the interaction between Ago2 and Let-7a, and distinguishing between strongly RNA-expressing cells and other cells, the interaction between Ago2 and Let-7a can be monitored dynamically, proving the utility of the probe. Probes will accelerate the basic role of reducing protein-RNA interactions, the mystery of RNA silencing, and monitoring the treatment of cancer.

我们精心设计了用于原位检测和成像Argonaute2（Ago2）的探针，Argonaute2（Ago2）是人细胞中的关键RNA干扰（RNAi）蛋白和siRNA（Let-7a）相互作用。该探针主要包含两部分：被淬灭的金纳米颗粒（AuNP）和作为荧光输出载体的功能化核酸。探针被内吞进入细胞后，修饰的发夹核酸可以被Ago2 / Let-7a复合物（ALC）切割成两部分。切割的核酸将替换在另一双链核酸上修饰的DNA链。替换的DNA链的3'末端被荧光团（Cy3）修饰。通过级联单个细胞，探针可以直接实现细胞中探针的荧光恢复。通过动态监控原位通过对Ago2和Let-7a之间相互作用的定量反应以及区分强表达RNA的细胞和其他细胞之间的相互作用，可以动态监控Ago2和Let-7a之间的相互作用，证明了该探针的实用性。探针将加速减少蛋白质-RNA相互作用，RNA沉默的奥秘以及监测癌症治疗的基本作用。