Leier André, Han Xu, Aghzadi Jehanne, Westin Erik, Liu Jian, Marquez Lago Tatiana T, Kesterson Robert A, Korf Bruce R, Wallis Deeann, Müller Ulrich F
Department of Genetics, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
Department of Chemistry & Biochemistry, University of California San Diego, La Jolla, CA 92093, USA.
Cancers (Basel). 2025 Aug 23;17(17):2749. doi: 10.3390/cancers17172749.
BACKGROUND/OBJECTIVES: Therapeutic strategies for Neurofibromatosis Type I (NF1) that correct the underlying pathogenic variant 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 from can be used to repair pathogenic variants of (pre-)mRNA by 3'-tail replacement.
Splice sites on the mRNA were identified computationally and validated biochemically, and an efficiency-enhancing Extended Guide Sequence (EGS) of the corresponding ribozyme was identified in a combinatorial experiment.
The correct trans-splicing product of this ribozyme was validated in HEK293 cells expressing .
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.
背景/目的：针对I型神经纤维瘤病（NF1）的治疗策略，若能纠正潜在的致病变异，则有望恢复神经纤维瘤蛋白功能、减轻肿瘤负担，并通过解决疾病的根本原因而非症状来改善患者预后。除了基因编辑，通过RNA反式剪接进行转录重编程也受到了关注，尤其是随着美国食品药品监督管理局（FDA）最近批准了两种基于反式剪接的药物进入1/2a期临床试验。本研究测试了来自[具体来源未提及]的反式剪接I组内含子核酶是否可用于通过3'-末端置换修复（前体）mRNA的致病变异。
通过计算确定了[基因名称未提及]mRNA上的剪接位点，并进行了生化验证，且在组合实验中确定了相应核酶的效率增强型扩展引导序列（EGS）。
在表达[相关蛋白未提及]的HEK293细胞中验证了该核酶的正确反式剪接产物。
本研究为NF1 mRNA的修复建立了剪接位点和活性增强型扩展引导序列。进一步优化核酶以及改进递送方法，可能会使基于核酶的RNA修复成为NF1治疗的可行策略。
