文章：

利用反式剪接I组内含子核酶修复突变的NF1 mRNA

Repair of MutatedNF1mRNA with Trans-Splicing Group I Intron Ribozymes

原文发布日期：23 August 2025

DOI: 10.3390/cancers17172749

类型: Article

开放获取: 是

英文摘要：

Background/Objectives: Therapeutic strategies for Neurofibromatosis Type I (NF1) that correct the underlying pathogenicNF1variant hold promise for restoring neurofibromin function, reducing tumor burden, and improving patient outcomes by addressing the root cause of the disease rather than its symptoms. Beyond gene editing, transcript reprogramming via RNA trans-splicing has gained attention, particularly with the recent FDA approval of two trans-splicing-based drugs for IND phase 1/2a trials. This study tests whether trans-splicing group I intron ribozymes fromTetrahymena thermophilacan be used to repair pathogenic variants ofNF1(pre-)mRNA by 3′-tail replacement. Methods: Splice sites on theNF1mRNA were identified computationally and validated biochemically, and an efficiency-enhancing Extended Guide Sequence (EGS) of the corresponding ribozyme was identified in a combinatorial experiment. Results: The correct trans-splicing product of this ribozyme was validated in HEK293NF1−/−cells expressingmNf1. Conclusions: This study established a splice site and activity-enhancing extended guide sequences for the repair of NF1 mRNA. Further optimization of the ribozyme, as well as improved delivery methods, may establish ribozyme-based RNA repair as a viable strategy for NF1 treatment.

摘要翻译： 

背景/目的：针对I型神经纤维瘤病（NF1）的治疗策略，通过纠正致病性NF1变异体，有望恢复神经纤维瘤蛋白功能、减轻肿瘤负荷，并通过解决疾病的根本原因而非症状来改善患者预后。除基因编辑外，通过RNA反式剪接进行的转录重编程已引起关注，特别是近期美国食品药品监督管理局批准了两种基于反式剪接的药物进入IND 1/2a期试验。本研究旨在验证嗜热四膜虫的I组内含子核酶是否可通过3'端替换修复NF1（前）mRNA的致病性变异。方法：通过计算分析识别NF1 mRNA上的剪接位点并进行生化验证，同时通过组合实验确定了相应核酶中增强效率的扩展引导序列。结果：该核酶在表达mNf1的HEK293NF1−/−细胞中验证了正确的反式剪接产物。结论：本研究确定了用于修复NF1 mRNA的剪接位点及增强活性的扩展引导序列。进一步优化核酶及改进递送方法，可能使基于核酶的RNA修复成为NF1治疗的可行策略。

原文链接：

Repair of MutatedNF1mRNA with Trans-Splicing Group I Intron Ribozymes