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末端维生素E修饰siRNA对基因表达的光调控
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摘要
设计并合成了用单个光活性基团和/或维生素E修饰在5'末端的干扰RNA。通过对细胞内萤火虫荧光素酶和绿色荧光蛋白这两种不同基因表达沉默的光调控的研究,发现这种设计的光敏感siRNA可实现其对基因表达的完全抑制和激活,并具有普适性。进一步的机制研究证明维生素E修饰的光敏感核酸与细胞表面的维生素E受体结合介导siRNA进入细胞,并与细胞内受体结合进一步增加位阻效应,从而实现高达18倍的光调控效果。
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 efficiency to date.siRNA tracking and vitE competition experiments indicated that the inactivity of vitE-modified siRNAs was not really 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.
引文
[1]Ji,Y.;Yang,J.;Wu,L.;Yu,L.;Tang,X.Angew.Chem.Int.Ed.2016,55,2152.

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