Elimination of Toxic Microsatellite Repeat Expansion RNA by RNA-Targeting Cas9.
Elimination of Toxic Microsatellite Repeat Expansion RNA by RNA-Targeting Cas9.
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DOI：
10.1016/j.cell.2017.07.010
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发表时间：
2017-08-24
期刊：
影响因子：
64.5
通讯作者：
Yeo GW
中科院分区：
文献类型：
--
作者：
Batra R;Nelles DA;Pirie E;Blue SM;Marina RJ;Wang H;Chaim IA;Thomas JD;Zhang N;Nguyen V;Aigner S;Markmiller S;Xia G;Corbett KD;Swanson MS;Yeo GW
DNA中的微卫星重复序列扩增产生致病的RNA物种，这些致病RNA物种主要导致遗传性疾病，如强直性肌营养不良1型和2型(DM1/2)、亨廷顿病和C9orf72连锁的肌萎缩侧索硬化症(C9-ALS)。为了诊断和治疗目的，需要以这些重复RNA为靶点的手段。在这里，我们描述了一种可编程的CRISPR系统的开发，该系统能够特定地可视化和消除这些有毒RNA。我们观察到当外源表达和在患者细胞中表达时，微卫星重复扩张RNA的特异性靶向和有效消除。重要的是，RNA靶向Cas9(RCas9)逆转了疾病的特征，包括消除了所有研究条件(DM1、DM2、C9-ALS、多谷氨酰胺疾病)中的RNA焦点，减少了多谷氨酰胺蛋白产物，重复结合蛋白的重新定位以类似于健康对照，以及有效地逆转了患者肌管中与DM1相关的剪接异常。最后，我们报告了一个与腺相关病毒包装兼容的截短RCas9系统。这一努力突显了RCas9在人类治疗方面的潜力。
Microsatellite repeat expansions in DNA produce pathogenic RNA species that cause dominantly inherited diseases such as myotonic dystrophy type 1 and 2 (DM1/2), Huntington’s disease, and C9orf72-linked amyotrophic lateral sclerosis (C9-ALS). Means to target these repetitive RNAs are required for diagnostic and therapeutic purposes. Here, we describe the development of a programmable CRISPR system capable of specifically visualizing and eliminating these toxic RNAs. We observe specific targeting and efficient elimination of micro-satellite repeat expansion RNAs both when exogenously expressed and in patient cells. Importantly, RNA-targeting Cas9 (RCas9) reverses hallmark features of disease including elimination of RNA foci among all conditions studied (DM1, DM2, C9-ALS, polyglutamine diseases), reduction of polyglutamine protein products, relocalization of repeat-bound proteins to resemble healthy controls, and efficient reversal of DM1-associated splicing abnormalities in patient myotubes. Finally, we report a truncated RCas9 system compatible with adeno-associated viral packaging. This effort highlights the potential of RCas9 for human therapeutics.
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影响因子：
4.5
作者：
Daughters RS;Tuttle DL;Gao W;Ikeda Y;Moseley ML;Ebner TJ;Swanson MS;Ranum LP
通讯作者：
Ranum LP
影响因子：
16.2
作者：
Ash PE;Bieniek KF;Gendron TF;Caulfield T;Lin WL;Dejesus-Hernandez M;van Blitterswijk MM;Jansen-West K;Paul JW 3rd;Rademakers R;Boylan KB;Dickson DW;Petrucelli L
通讯作者：
Petrucelli L
影响因子：
11.1
作者：
Lee, Kuang-Yung;Li, Moyi;Manchanda, Mini;Batra, Ranjan;Charizanis, Konstantinos;Mohan, Apoorva;Warren, Sonisha A.;Chamberlain, Christopher M.;Finn, Dustin;Hong, Hannah;Ashraf, Hassan;Kasahara, Hideko;Ranum, Laura P. W.;Swanson, Maurice S.
通讯作者：
Swanson, Maurice S.
影响因子：
11.2
作者：
Nakamori, Masayuki;Sobczak, Krzysztof;Puwanant, Araya;Welle, Steve;Eichinger, Katy;Pandya, Shree;Dekdebrun, Jeannne;Heatwole, Chad R.;McDermott, Michael P.;Chen, Tian;Cline, Melissa;Tawil, Rabi;Osborne, Robert J.;Wheeler, Thurman M.;Swanson, Maurice S.;Moxley, Richard T., III;Thornton, Charles A.
通讯作者：
Thornton, Charles A.
影响因子：
64.8
作者：
LASPADA, AR;WILSON, EM;FISCHBECK, KH
通讯作者：
FISCHBECK, KH