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.

RNA编辑技术，尤其是基于RECODE（条件稳定型增强ADAR1脱氨酶变体的RNA编辑技术），在精准表观转录组工程与治疗干预领域展现出巨大潜力。然而，转录组范围内的非预期脱靶修饰仍是其临床应用的主要顾虑。本项目旨在严格评估RECODE系统在HEK293T细胞中的全转录组特异性。我们对转染了RNA编辑元件的HEK293T细胞开展了高通量深度RNA测序（RNA-Seq），并以SYFP2等作为对照组进行比对分析。本研究的目标为鉴定并定量潜在的脱靶RNA编辑事件，评估其对全局基因表达谱的影响，并为该编辑平台提供全面的安全性特征图谱。本研究结果将为面向高保真应用的RNA编辑工具的后续优化提供有力支撑。