CRISPR-Cas13d targeting suppresses repeat-associated non-AUG translation of C9orf72 hexanucleotide repeat RNA.

Honghe Liu,Xiao-Feng Zhao,Yu-Ning Lu,Lindsey R Hayes,Jiou Wang
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Abstract

A hexanucleotide GGGGCC repeat expansion in the non-coding region of C9orf72 gene is the most common genetic mutation identified in patients with amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). The resulting repeat RNA and dipeptide repeat proteins from non-conventional repeat translation have been recognized as important markers associated with the diseases. CRISPR-Cas13d, a powerful RNA targeting tool, has faced challenges in effectively targeting RNA with stable secondary structures. Here we report that CRISPR-Cas13d can be optimized to specifically target GGGGCC repeat RNA. Our results demonstrate that the CRISPR-Cas13d system can be harnessed to significantly diminish the translation of poly-dipeptides originating from the GGGGCC repeat RNA. This efficacy has been validated in various cell types, including induced pluripotent stem cells and differentiated motor neurons originating from C9orf72-ALS patients, as well as in C9orf72 repeat transgenic mice. These findings demonstrate the application of CRISPR-Cas13d in targeting RNA with intricate higher-order structures and suggest a potential therapeutic approach for ALS and FTD.
CRISPR-Cas13d靶向抑制了C9orf72六核苷酸重复RNA的重复相关非AUG翻译。
C9orf72 基因非编码区的六核苷酸 GGGGCC 重复扩增是肌萎缩侧索硬化症(ALS)和额颞叶痴呆症(FTD)患者最常见的基因突变。非常规重复翻译产生的重复 RNA 和二肽重复蛋白被认为是与这些疾病相关的重要标志物。CRISPR-Cas13d 作为一种强大的 RNA 靶向工具,在有效靶向具有稳定二级结构的 RNA 方面一直面临挑战。在这里,我们报告了CRISPR-Cas13d可以优化为特异性靶向GGGGCC重复RNA。我们的研究结果表明,CRISPR-Cas13d 系统可以显著减少源自 GGGCC 重复 RNA 的多肽的翻译。这一功效已在多种细胞类型中得到验证,包括诱导多能干细胞、源自 C9orf72-ALS 患者的分化运动神经元以及 C9orf72 重复转基因小鼠。这些发现证明了CRISPR-Cas13d在靶向具有复杂高阶结构的RNA方面的应用,并为ALS和FTD提出了一种潜在的治疗方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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