Short-Chain Guide RNA for Site-Directed A-to-I RNA Editing
| Autor: | Masatora Fukuda, Kota Hidaka, Kanako Nose, Takashi Yano, Yohei Tomita |
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| Rok vydání: | 2021 |
| Předmět: |
Base Sequence
Adenosine Deaminase Oligonucleotide RNA-Binding Proteins RNA Computational biology Oligonucleotides Antisense Biology Biochemistry Target site RNA editing Drug Discovery ADAR RNA Sequence Genetics Humans Nucleic Acid Conformation Molecular Medicine Molecular Targeted Therapy RNA Editing RNA Messenger Guide RNA Molecular Biology RNA Guide Kinetoplastida |
| Zdroj: | Nucleic Acid Therapeutics. 31:58-67 |
| ISSN: | 2159-3345 2159-3337 |
| Popis: | Site-directed RNA editing is a promising genetic modification technology for therapeutic and pharmaceutical applications. We previously constructed adenosine deaminases acting on RNA (ADAR)-guiding RNAs (AD-gRNAs) that direct A-to-I RNA editing activity of native human ADAR2 into a programmable target site. In this study, we developed the short-chain AD-gRNA (shAD-gRNA) as a potential basic framework for practical RNA-editing oligonucleotides. Based on knowledge of previous AD-gRNA, shAD-gRNAs were designed to have the shortest possible sequence for the induction of editing activity. In vitro, compared to the original AD-gRNA, the shAD-gRNAs showed similar or superior editing induction activity, depending on the target RNA sequence, and had lower off-target editing activity around the target site, which is predicted to be a hotspot for off-target editing. Moreover, shAD-gRNAs achieved target RNA editing with both exogenous and endogenous human ADARs in cultured cells. Our results present shAD-gRNA as a short basic framework that would be applicable to further development for practical RNA-editing oligonucleotides. |
| Databáze: | OpenAIRE |
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