Repair of Mutated mRNA with Trans-Splicing Group I Intron Ribozymes.

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.

METHODS

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.

RESULTS

The correct trans-splicing product of this ribozyme was validated in HEK293 cells expressing .

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.