Accelerating Responsive RNA Release Through Structural Optimization of Disulfide‐Containing Acyl Groups

Abstract

Abstract Chemical methods for modifying and manipulating RNA are crucial for advancing its biological studies and applications. While postsynthetic 2′‐OH acylation has enabled on‐demand RNA activation, its application to larger, biologically relevant RNAs remains challenging. Herein, we present a redox‐responsive RNA modification via postsynthetic acylation to functionalize RNAs. Three strategies were developed to introduce multiple disulfide‐containing acyl adducts at 2′‐OH positions, temporarily blocking RNA function. Exposure to glutathione (GSH) can trigger the traceless release of RNA and restore its biological function. We demonstrate the versatility of this redox‐responsive strategy with RNA constructs of varying lengths, including short synthetic RNA, single guide RNA (sgRNA) as well as longer messenger RNA (mRNA). Furthermore, these disulfide‐containing acyl adducts respond to endogenous GSH, restoring mRNA translation without the need for cytotoxic exogenous stimuli. Taken together, these results offer a simple and generalizable method for modifying and modulating RNAs regardless of length or origin through structural optimization of acyl groups for facilitating RNA release, setting the stage for broad applicability.