State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences and Chemical Biology Center, Peking University, No. 38 Xueyuan Rd., Beijing 100191, China.
State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210023, China.
Molecules. 2022 Jul 8;27(14):4377. doi: 10.3390/molecules27144377.
The temporal activation of siRNA provides a valuable strategy for the regulation of siRNA activity and conditional gene silencing. The bioorthogonal bond-cleavage reaction of benzonorbonadiene and tetrazine is a promising trigger in siRNA temporal activation. Here, we developed a new method for the bio-orthogonal chemical activation of siRNA based on the tetrazine-induced bond-cleavage reaction. Small-molecule activatable caged siRNAs were developed with the 5'-vitamin E-benzonobonadiene-modified antisense strand targeting the green fluorescent protein (GFP) gene and the mitotic kinesin-5 (Eg5) gene. The addition of tetrazine triggered the reaction with benzonobonadiene linker and induced the linker cleavage to release the active siRNA. Additionally, the conditional gene silencing of both exogenous GFP and endogenous Eg5 genes was successfully achieved with 5'-vitamin E-benzonobonadiene-caged siRNAs, which provides a new uncaging strategy with small molecules.
siRNA 的时空调控为调控 siRNA 活性和条件性基因沉默提供了一种有价值的策略。苯并降蒈二烯和四嗪的生物正交键断裂反应是 siRNA 时空调控的一种很有前途的触发机制。在这里，我们基于四嗪诱导的键断裂反应，开发了一种新的 siRNA 生物正交化学激活方法。设计了带有 5'-生育酚-苯并降蒈二烯修饰的反义链的小分子可激活的环化 siRNA，该反义链靶向绿色荧光蛋白 (GFP) 基因和有丝分裂驱动蛋白-5 (Eg5) 基因。四嗪的加入引发了与苯并降蒈二烯连接子的反应，并诱导连接子断裂释放活性 siRNA。此外，用 5'-生育酚-苯并降蒈二烯环化 siRNA 成功实现了外源性 GFP 和内源性 Eg5 基因的条件性基因沉默，为小分子提供了一种新的解笼策略。
