Nitro Reduction-Based RNA Control and Ultrafast Release

Abstract

RNA protection and controlled release are critical for both fundamental research and therapeutic applications, yet the development of simple, efficient, and reversible post-synthetic RNA modification strategies remains a significant challenge. Here, we introduce a straight forward approach based on ribose 2'-hydroxyl acylation with a nitro-functionalized carbamate, which acts as a redox-responsive center. This modification selectively inhibits native RNA function while remaining inert to endogenous biogenic reductants and common reducing agents used in biological assays. Upon treatment with low millimolar concentrations of the THDB- BIPY reducing pair, RNA function is rapidly restored on a minute timescale. This methodology is broadly applicable across diverse RNA classes and functional contexts, including synthetic RNA oligomers, fluorogenic RNA aptamers, single-guide RNAs in CRISPR-Cas9 gene-editing systems, and mRNAs in living cells for translation control and RNAi-mediated gene silencing. These results demonstrate the general utility of this approach as a chemically controllable functional switch, providing a versatile toolkit for temporal regulation of RNA activity in both research and biotechnological applications.