Department of Chemistry, Stanford University, Stanford, CA 94305, USA.
Org Biomol Chem. 2021 Oct 6;19(38):8367-8376. doi: 10.1039/d1ob01713f.
Caging RNA by polyacylation (cloaking) has been developed recently as a simple and rapid method to control the function of RNAs. Previous approaches for chemical reversal of acylation (uncloaking) made use of azide reduction followed by amine cyclization, requiring ∼2-4 h for the completion of cyclization. In new studies aimed at improving reversal rates and yields, we have designed novel acylating reagents that utilize quinone methide (QM) elimination for reversal. The QM de-acylation reactions were tested with two bioorthogonally cleavable motifs, azide and vinyl ether, and their acylation and reversal efficiencies were assessed with NMR and mass spectrometry on model small-molecule substrates as well as on RNAs. Successful reversal both with phosphines and strained alkenes was documented. Among the compounds tested, the azido-QM compound A-3 displayed excellent de-acylation efficiency, with for de-acylation of less than an hour using a phosphine trigger. To test its function in RNA caging, A-3 was successfully applied to control EGFP mRNA translation and in HeLa cells. We expect that this molecular caging strategy can serve as a valuable tool for biological investigation and control of RNAs both and in cells.
聚酰化（伪装）将 RNA 笼闭最近被开发为一种简单快速的控制 RNA 功能的方法。先前用于化学反转酰化（解伪装）的方法利用叠氮还原 followed by amine cyclization，需要约 2-4 h 完成环化。在旨在提高反转速率和产率的新研究中，我们设计了利用醌甲醚 (QM) 消除进行反转的新型酰化试剂。QM 去酰化反应用两种生物正交可裂解的基序（叠氮化物和乙烯基醚）进行了测试，并用 NMR 和质谱对小分子模型底物以及 RNA 上的酰化和反转效率进行了评估。成功地用膦和张力烯烃进行了反转。在所测试的化合物中，叠氮-QM 化合物 A-3 显示出极好的去酰化效率，不到一小时的时间内使用膦触发剂即可达到 。为了测试其在 RNA 笼闭中的功能，A-3 成功地应用于控制 EGFP mRNA 翻译和 HeLa 细胞。我们预计，这种分子笼闭策略可以作为生物研究和控制 RNA 的有价值的工具，无论是在体外还是在细胞内。
