Adenine Base Editor Ribonucleoproteins Delivered by Lentivirus-Like Particles Show High On-Target Base Editing and Undetectable RNA Off-Target Activities
| Autor: | Baisong Lu, Kyung Whan Yoo, Sung-Ik Cho, Anthony Atala, Zuyan Lu, Manish Kumar Yadav, Jin-Soo Kim, Pin Lyu |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Aptamer
Gene mutation Capsid Genetics Humans Ribonucleoprotein Gene Editing biology Base Sequence Chemistry Electroporation Adenine Lentivirus RNA Aptamers Nucleotide biology.organism_classification Cell biology HEK293 Cells Ribonucleoproteins RNA editing Mutation RNA Editing Biotechnology RNA Guide Kinetoplastida |
| Zdroj: | The CRISPR journal. 4(1) |
| ISSN: | 2573-1602 |
| Popis: | Adenine base editors (ABEs) can correct gene mutations without creating double-strand breaks. However, in recent reports, these editors showed guide-independent RNA off-target activities. This work describes our development of a delivery method to minimize ABEs' RNA off-target activity. After discovering a RNA off-target hot spot for sensitive detection of RNA off-target activities, we found that delivering ribonucleoproteins (RNPs) by electroporation generated undetectable non-specific RNA editing, but on-target base editing activity was also relatively low. We then explored a lentivirus capsid-based delivery strategy to deliver ABE. We used aptamer/aptamer-binding protein (ABP) interactions to package ABE RNPs into lentiviral capsids. Capsid RNPs were delivered to human cells for highly efficient guided base editing. Importantly, RNA off-target activities from the capsid RNPs were undetectable. Our new lentiviral capsid-based ABE RNP delivery method with minimal RNA off-target activities makes ABE one step closer to possible therapeutic applications. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|