Switchable genome editing via genetic code expansion
Switchable genome editing via genetic code expansion
复制标题
通过遗传密码扩展进行可切换的基因组编辑
DOI：
10.1101/349951
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Suzuki T
中科院分区：
文献类型：
--
作者：
Suzuki T
CRISPR-Cas9基因组编辑的多种应用需要严格的调控，因此产生了Cas9变体，其活性对小配体、温度或光有响应。然而，这些方法通常是不切实际的，例如在临床治疗性基因组编辑或基因驱动中，环境相容控制是至关重要的。考虑到这一点，我们开发了可遗传的cas9介导的哺乳动物基因组编辑，该编辑由廉价的赖氨酸衍生物Lys(Boc) （Boc）严格控制。遗传密码扩增允许非生理性的BOC掺入，使得Cas9 （Cas9BOC）仅在BOC暴露后才能在培养的体细胞中以全长、活性形式表达。当在小鼠胚胎发育开始时，从cRNA或cas9boc转基因雌性中表达cas9boc时，发生了严格依赖于boc的转基因和原生基因组位点的可遗传编辑。这里报道的严格控制的Cas9编辑系统有望有广泛的应用，是在大地理范围内有目的、时空基因驱动调控的第一步。
Multiple applications of genome editing by CRISPR-Cas9 necessitate stringent regulation and Cas9 variants have accordingly been generated whose activity responds to small ligands, temperature or light. However, these approaches are often impracticable, for example in clinical therapeutic genome editingin situor gene drives in which environmentally-compatible control is paramount. With this in mind, we have developed heritable Cas9-mediated mammalian genome editing that is acutely controlled by the cheap lysine derivative, Lys(Boc) (BOC). Genetic code expansion permitted non-physiological BOC incorporation such that Cas9 (Cas9BOC) was expressed in a full-length, active form in cultured somatic cells only after BOC exposure. Stringently BOC-dependent, heritable editing of transgenic and native genomic loci occurred when Cas9BOCwas expressed at the onset of mouse embryonic development from cRNA orCas9BOCtransgenic females. The tightly controlled Cas9 editing system reported here promises to have broad applications and is a first step towards purposed, spatiotemporal gene drive regulation over large geographical ranges.
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