A high-fidelity RNA-targeting Cas13 restores paternal Ube3a expression and improves motor functions in Angelman syndrome mice
Brief intro:
- Author: Jinhui Li, Zhixin Shen, Yajing Liu, Zixiang Yan, Yuanhua Liu, Xiang Lin, Junjie Tang, Ruimin Lv, Guannan Geng, Zhi-Qi Xiong, Changyang Zhou, Hui Yang
- Journal: Molecular Therapy
- Doi: https://www.doi.org/10.1016/j.ymthe.2023.02.015
- Publication Date: 2023 Feb 18
Products/Services used in the paper
Quotation shows PackGene:AAV-PHP.eb serotype was used in this study. The CAG-tdTomato,hSyn1-hfCas13x.1/U6-cr9, or hfCas13x.1/U6-NT plasmids were constructed and sequenced before packaging into AAV-PHP.eb vehicle; and the AAV vectors were packaged by PackGene Biotech. The vector titer was determined by qPCR specific for the inverted terminal repeat.
Research Field:CNS
AAV Serotype:AAV-PHP.eb
Targeted organ:brain
Animal or cell line strain:Ube3am-/p+ mice and Ube3am+/pYFP mice were maintained on the C57Bl/6 background.
Abstract
Angelman syndrome (AS) is a rare neurodevelopmental disorder caused by loss of function mutations in maternally expressed UBE3A. No gene-specific treatment is available for patients so far. Although intact and transcriptionally active, paternally inherited UBE3A is silenced by elongation of antisense long noncoding RNA UBE3A-ATS in neurons. Here, we demonstrated that RNA targeting of paternal Ube3a-ATS with a high-fidelity CRISPR-Cas13 (hfCas13x.1) system could restore Ube3a expression to similar levels as that of maternal Ube3a in the cultured mouse neurons. Furthermore, injection into lateral ventricles with neuron-specific hSyn1 promoter-driven hfCas13x.1 packaged in adeno-associated virus (AAV-PHP.eb) could restore paternal Ube3a expression in cortex and hippocampus of neonatal AS mice for up to 4 months after treatment. Behavioral tests showed that expression of paternal Ube3a significantly alleviated AS-related symptoms, including obesity and motor function. Our results suggested that hfCas13x.1-mediated suppression of the Ube3a-ATS lncRNA potentially serves as a promising targeted intervention for AS.
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