Design and Validation of Mirtron-Mediated RNA Interference for Gene Therapy.

Zheng Q., Orlans HO., Staurenghi F., McClements ME., MacLaren RE.

Mirtrons, a subclass of intronic microRNAs, offer an alternative RNA interference (RNAi) mechanism by bypassing Drosha processing. This protocol details the design and production of mirtron-based constructs aimed at achieving efficient gene knockdown. By embedding mirtrons in the 5' untranslated region (UTR) of a transgene, we evaluated processing efficiency and RNAi activity. The compact size of mirtrons facilitates concurrent gene knockdown and supplementation within a single adeno-associated virus (AAV) vector. Our findings present a novel strategy for gene regulation in therapeutic and research applications.

DOI

10.1007/978-1-0716-5198-8_14

Type

Journal article

Publication Date

2026-01-01T00:00:00+00:00

Volume

3023

Pages

195 - 206

Total pages

11

Keywords

DNA cloning, Gene therapy, Mirtron, RNAi, MicroRNAs, RNA Interference, Dependovirus, Genetic Therapy, Humans, Genetic Vectors, 5' Untranslated Regions, Gene Knockdown Techniques, Introns

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