Photoinduced RNA interference using DMNPE-caged 2′-deoxy-2′-fluoro substituted nucleic acids in vitro and in vivo
Photoinduced RNA interference using DMNPE-caged 2′-deoxy-2′-fluoro substituted nucleic acids in vitro and in vivo
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DOI：
10.1039/b801532e
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发表时间：
2008-01-01
影响因子：
--
通讯作者：
Monroe, W. Todd
中科院分区：
文献类型：
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作者：
Blidner, Richard A.;Svoboda, Kurt R.;Monroe, W. Todd
各种化学物质，莫迪。阳离子被加入到RNA中，试图改善它们对RNAi干扰(RNAi)的药理性质。最近的研究表明，含有2‘-氟代Modi的小干扰RNA(SiRNA)。阳离子可通过RNAi诱导基因沉默。尽管在使用化学物质莫迪方面取得了进展。尽管阳离子药物能够提高这些疗法的稳定性、安全性和有效性，但它们仍然面临着空间和时间靶向的挑战。一种潜在的靶向策略是使用光笼技术，这涉及到耐光性化合物与效应器核酸物种的共价连接，这些核酸物种阻止生物活性，直到暴露在近紫外光下。在这项研究中，我们证明了在细胞培养和斑马鱼胚胎中，完全的2‘-氟化核酸(FNAs)可以被关在笼子里用于光激活的基因沉默。这一策略结合了与2‘-取代相关的化学和酶稳定性的改善与光诱导触发的靶向能力。FNAs与1-(4，5-二甲氧基-2-硝基苯基)重氮乙烷(DMNPE)的统计烷基化反应提高了对酶降解的抵抗力，降低了RNAi效果，并保护了生物系统免受效应器的毒性剂量的影响。365 nm光照射部分恢复了2‘-氟代siRNA的沉默活性。这些结果表明，光笼化可能超过了对RNAi疗法的空间和时间定向激活的控制，同时提高了酶的稳定性，并可能通过光剂量强度实现治疗性剂量。
Various chemical modi. cations to RNA have been incorporated in attempts to improve their pharmacological properties for RNAi interference (RNAi). Recent studies have shown that small interfering RNA (siRNA) containing 2'-fluoro modi. cations can elicit gene silencing through RNAi. Despite developments in using chemical modi. cations for increased stability, safety, and efficiency of these therapeutics, they still face challenges of spatial and temporal targeting. One potential targeting strategy is to use photocaging techniques, which involve the covalent attachment of photolabile compounds to the effector nucleic acid species that block bioactivity until exposed to near UV light. In this study we demonstrate that fully 2'-fluorinated nucleic acids (FNAs) can be caged for photoactivated gene silencing in cell culture and in zebrafish embryos. This strategy combines the improvement in chemical and enzymatic stability associated with 2'-substitutions with the targeting ability of a photoinducible trigger. Statistical alkylation of FNAs with 1-(4,5-dimethoxy-2-nitrophenyl) diazoethane (DMNPE) improved resistance to enzymatic degradation, reduced RNAi effectiveness, and protected the biological system from toxic doses of the effector. Photo-exposure to 365 nm light partially restored the silencing activity of the 2'-fluoro siRNAs. These results suggest that photocaging may over control over RNAi therapeutics for spatially and temporally directed activation, while improving enzymatic stability and potentially enabling therapeutic dosing via light dose intensity.