State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, No. 38, Xueyuan Rd., Beijing, 100191, China.
Angew Chem Int Ed Engl. 2016 Feb 5;55(6):2152-6. doi: 10.1002/anie.201510921. Epub 2015 Dec 28.
Caged siRNAs with a single photolabile linker and/or vitamin E (vitE) modification at the 5' terminal were rationally designed and synthesized. These virtually inactive caged siRNAs were successfully used to photoregulate both firefly luciferase and GFP gene expression in cells with up to an 18.6-fold enhancement of gene silencing activity, which represents one of the best reported photomodulation of gene silencing efficiencies to date. siRNA tracking and vitE competition experiments indicated that the inactivity of vitE-modified siRNAs was not due to the bulky moiety of vitE; rather, the involvement of vitE-binding proteins has a large contribution to caged siRNA inactivation by preventing the dissociation of siRNA/lipo complexes and/or siRNA release. Further patterning experiments revealed the ability to spatially regulate gene expression through simple light irradiation.
设计并合成了带有单个光解连接子和/或 5' 端维生素 E（vitE）修饰的笼状 siRNA。这些几乎无活性的笼状 siRNA 成功地用于光调控细胞内萤火虫荧光素酶和 GFP 基因的表达，基因沉默活性增强高达 18.6 倍，这是迄今为止报道的最好的光调控基因沉默效率之一。siRNA 追踪和 vitE 竞争实验表明，vitE 修饰的 siRNA 的无活性不是由于 vitE 的庞大部分；相反，vitE 结合蛋白的参与通过阻止 siRNA/脂质复合物的解离和/或 siRNA 释放对笼状 siRNA 的失活有很大贡献。进一步的图案化实验表明，通过简单的光照射能够实现空间调节基因表达。
