Abstract
RNA editing is a prospective therapeutic approach for correcting harmful mutations, offering the benefits of reversibility and tunability without permanently modifying the genome. However, the relatively low enzymatic activity and the occurrence of off-target editing events present significant challenges, limiting its utility. In response to this limitation, we introduced a novel strategy: strand displacement-responsive ADAR system for RNA editing (SPRING) by adding a "blocking sequence" to form a hairpin guide RNA. This modification significantly improves the efficiency of site-directed RNA editing (SDRE) at various target sites. Furthermore, the use of hairpin guide RNA within the SPRING system enhances the specificity of RNA editing through competitive reactions during target hybridization. In principle, this approach can be employed across various ADAR-based editing systems, offering a novel RNA-editing platform with wide-ranging potential for research, therapy, and biotech applications.