持续表达靶向毒性 RNA 的 Cas9 可逆转肌强直性营养不良 1 型小鼠模型的疾病表型。
The sustained expression of Cas9 targeting toxic RNAs reverses disease phenotypes in mouse models of myotonic dystrophy type 1.
机构信息
Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, CA, USA.
Stem Cell Program, University of California San Diego, La Jolla, CA, USA.
出版信息
Nat Biomed Eng. 2021 Feb;5(2):157-168. doi: 10.1038/s41551-020-00607-7. Epub 2020 Sep 14.
Myotonic dystrophy type I (DM1) is a multisystemic autosomal-dominant inherited human disorder that is caused by CTG microsatellite repeat expansions (MREs) in the 3' untranslated region of DMPK. Toxic RNAs expressed from such repetitive sequences can be eliminated using CRISPR-mediated RNA targeting, yet evidence of its in vivo efficacy and durability is lacking. Here, using adult and neonatal mouse models of DM1, we show that intramuscular or systemic injections of adeno-associated virus (AAV) vectors encoding nuclease-dead Cas9 and a single-guide RNA targeting CUG repeats results in the expression of the RNA-targeting Cas9 for up to three months, redistribution of the RNA-splicing protein muscleblind-like splicing regulator 1, elimination of foci of toxic RNA, reversal of splicing biomarkers and amelioration of myotonia. The sustained reversal of DM1 phenotypes provides further support that RNA-targeting Cas9 is a viable strategy for treating DM1 and other MRE-associated diseases.
Ⅰ型肌强直性营养不良（DM1）是一种多系统常染色体显性遗传的人类疾病，由 DMPK 3'非翻译区的 CTG 微卫星重复扩展（MREs）引起。可以使用 CRISPR 介导的 RNA 靶向来消除来自这种重复序列的毒性 RNA，但缺乏其体内疗效和持久性的证据。在这里，我们使用 DM1 的成年和新生小鼠模型表明，肌内或系统注射编码核酸酶失活 Cas9 和靶向 CUG 重复的单链引导 RNA 的腺相关病毒（AAV）载体可使 RNA 靶向 Cas9 的表达长达三个月，RNA 剪接蛋白肌肉盲样剪接调节剂 1 的重新分布，消除毒性 RNA 的焦点，逆转剪接生物标志物，并改善肌强直。DM1 表型的持续逆转进一步支持 RNA 靶向 Cas9 是治疗 DM1 和其他 MRE 相关疾病的可行策略。
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