Engineered circular ADAR-recruiting RNAs increase the efficiency and fidelity of RNA editing in vitro and in vivo
简介:
- 作者: 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
- 杂志: Nature Biotechnology
- Doi: https://www.doi.org/10.1038/s41587-021-01180-3
- 出版日期: 2022 Jun
论文中使用的产品/服务
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).
摘要
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.
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作为一家专注于AAV 技术十余年，深耕基因治疗领域的CRO&CDMO，派真生物可提供从载体设计、构建到 AAV、慢病毒和 mRNA 服务的一站式解决方案。凭借深厚的技术实力、卓越的运营管理和高标准的服务交付，我们为全球客户提供一站式CMC解决方案，包括从早期概念验证、成药性评估到IIT、IND及BLA的各个阶段。
凭借我们独立知识产权的π-alphaTM 293 细胞AAV高产技术平台，我们能将AAV产量提高多至10倍，每批次产量可达1×10¹⁷vg，以满足多样化的商业化和临床项目需求。此外，我们定制化的mRNA和脂质纳米颗粒（LNP）产品及服务覆盖药物和疫苗开发的各个阶段，从研发到符合GMP的生产，提供端到端的一站式解决方案。
