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

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.