Evaluation of the specificity and global off-target effects of a novel RNA editing tool RECODE via RNA-Seq
RNA editing technologies, particularly those based on the RECODE (RNA editing with conditionally stable and enhanced ADARl deaminasevariants), offer great potential for precise epitranscriptome engineering and therapeutic intervention. However, unintended off-target modifications across the transcriptome remain a major concern for their clinical application. This project aims to rigorously evaluate the transcriptome-wide specificity of the RECODE system in HEK293T cells. We performed high-depth RNA sequencing (RNA-Seq) on HEK293T cells transfected with the RNA editing components and compared them with control groups (e.g., SYFP2). The goal of this study is to identify and quantify potential off-target RNA editing events, assess the impact on the global gene expression profile, and provide a comprehensive safety profile for this editing platform. The findings will support the further optimization of RNA editing tools for high-fidelity applications.