Nature Biotechnology. 2022 Jun
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 
Products used in the paper Details Operation
AAV vector packaging 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). Request Quote

Research Field: RNA editing

AAV Serotype: AAV8

Dose: 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).

Routes of Administration: tail vein

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.

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