通过 CRISPR-Cas9 靶向细胞 mRNA 翻译。
Targeting cellular mRNAs translation by CRISPR-Cas9.
机构信息
Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen 518039, China.
Department of Urological Surgery, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen 518039, China.
出版信息
Sci Rep. 2016 Jul 13;6:29652. doi: 10.1038/srep29652.
Recently CRISPR-Cas9 system has been reported to be capable of targeting a viral RNA, and this phenomenon thus raises an interesting question of whether Cas9 can also influence translation of cellular mRNAs. Here, we show that both natural and catalytically dead Cas9 can repress mRNA translation of cellular genes, and that only the first 14 nt in the 5' end of sgRNA is essential for this process. CRISPR-Cas9 can suppress the protein expression of an unintended target gene without affecting its DNA sequence and causes unexpected phenotypic changes. Using the designed RNA aptamer-ligand complexes which physically obstruct translation machinery, we indicate that roadblock mechanism is responsible for this phenomenon. Our work suggests that studies on Cas9 should avoid the potential off-target effects by detecting the alteration of genes at both the DNA and protein levels.
最近有报道称 CRISPR-Cas9 系统能够靶向病毒 RNA，这就提出了一个有趣的问题，即 Cas9 是否也能影响细胞 mRNA 的翻译。在这里，我们表明，天然 Cas9 和无酶活性的 Cas9 都能抑制细胞基因的 mRNA 翻译，而 sgRNA 5' 端的前 14 个核苷酸对于这一过程是必需的。CRISPR-Cas9 可以在不影响其 DNA 序列的情况下抑制非预期靶标基因的蛋白质表达，并导致意外的表型变化。使用设计的 RNA 适体-配体复合物来物理阻断翻译机制，我们表明，这种现象是由阻碍机制引起的。我们的工作表明，Cas9 的研究应该通过检测 DNA 和蛋白质水平上基因的变化来避免潜在的脱靶效应。
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