小柯机器人

Cas9实现基因治疗Angelman综合征
2020-10-23 14:36

美国北卡罗莱纳大学教堂山分校Mark J. Zylka小组利用Cas9实现基因治疗Angelman综合征。相关论文于2020年10月21日在线发表于国际学术期刊《自然》。

据研究人员介绍,Angelman综合征(AS)是由母亲遗传的UBE3A等位基因突变或缺失引起的严重神经发育障碍。在神经元中,父系继承的UBE3A等位基因被称为UBE3A-ATS的长非编码RNA顺式沉默。

作为系统筛选的一部分,研究人员发现当靶向Snord115基因(聚集在Ube3a-ATS 3'区的小核仁RNA)时,Cas9可用于激活培养的小鼠和人类神经元中的父系Ube3a(“去沉默”)。靶向约75个Snord115基因的短Cas9变体和向导RNA被包装到腺相关病毒中,并在胚胎和出生后早期阶段被应用于AS的小鼠模型,预计恢复Ube3a的治疗益处将会最大。这种早期治疗使整个父亲的父亲Ube3a沉默了至少17个月,并挽救了AS小鼠的解剖学和行为表型。腺相关病毒载体基因组整合到Cas9目标位点中,导致Ube3a-ATS在载体来源的polyA盒中提前终止,或者在反向整合时,通过与载体衍生的Cas9转录物的转录碰撞而终止。

这项研究表明,基因治疗载体的靶向基因组整合可以在整个生命中恢复父系继承的UBE3A的功能,为综合征神经发育障碍的疾病缓解提供了途径。

附:英文原文

Title: Cas9 gene therapy for Angelman syndrome traps Ube3a-ATS long non-coding RNA

Author: Justin M. Wolter, Hanqian Mao, Giulia Fragola, Jeremy M. Simon, James L. Krantz, Hannah O. Bazick, Baris Oztemiz, Jason L. Stein, Mark J. Zylka

Issue&Volume: 2020-10-21

Abstract: Angelman syndrome (AS) is a severe neurodevelopmental disorder caused by a mutation or deletion of the maternally inherited UBE3A allele. In neurons, the paternally inherited UBE3A allele is silenced in cis by a long non-coding RNA called UBE3A-ATS. Here, as part of a systematic screen, we found that Cas9 can be used to activate ('unsilence') paternal Ube3a in cultured mouse and human neurons when targeted to Snord115 genes, which are small nucleolar RNAs that are clustered in the 3′ region of Ube3a-ATS. A short Cas9 variant and guide RNA that target about 75 Snord115 genes were packaged into an adeno-associated virus and administered to a mouse model of AS during the embryonic and early postnatal stages, when the therapeutic benefit of restoring Ube3a is predicted to be greatest1,2. This early treatment unsilenced paternal Ube3a throughout the brain for at least 17 months and rescued anatomical and behavioural phenotypes in AS mice. Genomic integration of the adeno-associated virus vector into Cas9 target sites caused premature termination of Ube3a-ATS at the vector-derived polyA cassette, or when integrated in the reverse orientation, by transcriptional collision with the vector-derived Cas9 transcript. Our study shows that targeted genomic integration of a gene therapy vector can restore the function of paternally inherited UBE3A throughout life, providing a path towards a disease-modifying treatment for a syndromic neurodevelopmental disorder.

DOI: 10.1038/s41586-020-2835-2

Source: https://www.nature.com/articles/s41586-020-2835-2

Nature:《自然》,创刊于1869年。隶属于施普林格·自然出版集团,最新IF:43.07
官方网址:http://www.nature.com/
投稿链接:http://www.nature.com/authors/submit_manuscript.html


本期文章:《自然》:Online/在线发表

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