Engineered circular ADAR-recruiting RNAs increase the efficiency and fidelity of RNA editing in vitro and in vivo
Brief intro:
- Author: Zongyi Yi , Liang Qu , Huixian Tang, Zhiheng Liu, Ying Liu, Feng Tian , Chunhui Wang, Xiaoxue Zhang, Ziqi Feng, Ying Yu, Pengfei Yuan, Zexuan Yi, Yanxia Zhao and Wensheng Wei
- Journal: Nature Biotechnology
- Doi: https://www.doi.org/10.1038/s41587-021-01180-3
- Publication Date: 2022 Jun
Products/Services used in the paper
Quotation shows PackGene:Circ-arRNAs were packaged in AAV8 by PackGene Biotech. The AAV titer was 1 × 10^13 virus/200 μl; 200 μl of AAV was injected into the tail vein of each IDUA-W392X mouse. Mice were monitored four times per week for the duration of the experiment (4 weeks).
Research Field:RNA editing
AAV Serotype:AAV8
Animal or cell line strain:The experimental animals included 4- or 6-week-old Idua-W392X (B6.129S-Iduatm1.1Kmke/J) female mice (Jackson Laboratory, no. 017681) and C57BL/6 J female mice (Beijing Vital River Laboratory).
Abstract
Current methods for programmed RNA editing using endogenous ADAR enzymes and engineered ADAR-recruiting RNAs (arRNAs) suffer from low efficiency and bystander off-target editing. Here, we describe LEAPER 2.0, an updated version of LEAPER that uses covalently closed circular arRNAs, termed circ-arRNAs. We demonstrate on average ~3.1-fold higher editing efficiency than their linear counterparts when expressed in cells or delivered as in vitro-transcribed circular RNA oligonucleotides. To lower off-target editing we deleted pairings of uridines with off-target adenosines, which almost completely eliminated bystander off-target adenosine editing. Engineered circ-arRNAs enhanced the efficiency and fidelity of editing endogenous CTNNB1 and mutant TP53 transcripts in cell culture. Delivery of circ-arRNAs using adeno-associated virus in a mouse model of Hurler syndrome corrected the pathogenic point mutation and restored α-L-iduronidase catalytic activity, lowering glycosaminoglycan accumulation in the liver. LEAPER 2.0 provides a new design of arRNA that enables more precise, efficient RNA editing with broad applicability for therapy and basic research.
About PackGene
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