利用 rAAV 载体在小鼠中实现 microRNA 功能的长期、高效抑制。
Long-term, efficient inhibition of microRNA function in mice using rAAV vectors.
机构信息
Gene Therapy Center, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
出版信息
Nat Methods. 2012 Mar 4;9(4):403-9. doi: 10.1038/nmeth.1903.
Understanding the function of individual microRNA (miRNA) species in mice would require the production of hundreds of loss-of-function strains. To accelerate analysis of miRNA biology in mammals, we combined recombinant adeno-associated virus (rAAV) vectors with miRNA 'tough decoys' (TuDs) to inhibit specific miRNAs. Intravenous injection of rAAV9 expressing anti-miR-122 or anti-let-7 TuDs depleted the corresponding miRNA and increased its mRNA targets. rAAV producing anti-miR-122 TuD but not anti-let-7 TuD reduced serum cholesterol by >30% for 25 weeks in wild-type mice. High-throughput sequencing of liver miRNAs from the treated mice confirmed that the targeted miRNAs were depleted and revealed that TuDs induced miRNA tailing and trimming in vivo. rAAV-mediated miRNA inhibition thus provides a simple way to study miRNA function in adult mammals and a potential therapy for dyslipidemia and other diseases caused by miRNA deregulation.
理解小鼠中个别 microRNA（miRNA）种类的功能需要产生数百种功能丧失株。为了加速哺乳动物中 miRNA 生物学的分析，我们将重组腺相关病毒（rAAV）载体与 miRNA“坚韧的诱饵”（TuD）相结合，以抑制特定的 miRNA。表达抗 miR-122 或抗 let-7 TuD 的 rAAV9 的静脉内注射耗尽了相应的 miRNA 并增加了其 mRNA 靶标。在 25 周内，rAAV 产生抗 miR-122 TuD 但不产生抗 let-7 TuD 的野生型小鼠血清胆固醇降低了 >30%。用处理后的小鼠的肝脏 miRNA 的高通量测序证实了靶向 miRNA 被耗尽，并表明 TuD 在体内诱导了 miRNA 的加尾和修剪。rAAV 介导的 miRNA 抑制因此为研究成年哺乳动物中的 miRNA 功能提供了一种简单的方法，并且为 miRNA 失调引起的血脂异常和其他疾病提供了一种潜在的治疗方法。
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