Department of Chemistry, The Scripps Research Institute, Scripps Florida, 130 Scripps Way, 3A1, Jupiter, FL 33458, USA.
Angew Chem Int Ed Engl. 2013 Jan 28;52(5):1462-5. doi: 10.1002/anie.201206888. Epub 2012 Dec 20.
Antisense oligonucleotides and small interfering RNAs (siRNAs) control gene expression by triggering the degradation of a mRNA via recruitment of RNase H or the RNA-induced silencing complex (RISC), respectively. These approaches are hampered, however, by the poor cellular permeability of oligonucleotides. A small molecule approach to cleave RNA targets could obviate uptake issues. Several compounds can induce RNA cleavage , however, to the best of our knowledge no small molecules have been previously described to cleave RNA in living cells. Herein, we describe the development of a potentially general approach to design small molecules that specifically cleave an RNA in a living cell, affecting biological function. Specifically, a designed, modularly assembled small molecule that binds the RNA that causes myotonic dystrophy type 1 (DM1) was appended with a moiety that generates hydroxyl radicals upon irradiation. Cleavage of the transcript improves DM1-associated defects in cell culture, and compounds are non-toxic at an efficacious dose as determined by a MTT viability assay. This approach may allow for the site-specific cleavage and inactivation of other cellular RNAs. Compounds that bind to and cleave RNA have the potential to serve as chemical genetics probes of function or lead therapeutics with spatial and temporal control.
反义寡核苷酸和小干扰 RNA（siRNA）通过分别招募核糖核酸酶 H 或 RNA 诱导沉默复合物（RISC）来触发 mRNA 的降解，从而控制基因表达。然而，这些方法受到寡核苷酸细胞通透性差的阻碍。一种切割 RNA 靶标的小分子方法可以避免摄取问题。有几种化合物可以诱导 RNA 切割，但是据我们所知，以前没有描述过小分子可以在活细胞中切割 RNA 以影响生物功能。在此，我们描述了一种设计专门在活细胞中切割特定 RNA 的小分子的潜在通用方法，从而影响生物功能。具体而言，设计了一种模块化组装的小分子，该小分子与导致 1 型肌强直性营养不良（DM1）的 RNA 结合，并附加了一个在照射时产生羟基自由基的部分。该转录物的切割改善了细胞培养中与 DM1 相关的缺陷，并且通过 MTT 活力测定确定，在有效剂量下化合物没有毒性。这种方法可能允许对其他细胞 RNA 进行特异性切割和失活。与 RNA 结合并切割 RNA 的化合物具有作为化学遗传学功能探针或具有时空控制的治疗剂的潜力。
