Yin Hao, Song Chun-Qing, Dorkin Joseph R, Zhu Lihua J, Li Yingxiang, Wu Qiongqiong, Park Angela, Yang Junghoon, Suresh Sneha, Bizhanova Aizhan, Gupta Ankit, Bolukbasi Mehmet F, Walsh Stephen, Bogorad Roman L, Gao Guangping, Weng Zhiping, Dong Yizhou, Koteliansky Victor, Wolfe Scot A, Langer Robert, Xue Wen, Anderson Daniel G
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
Nat Biotechnol. 2016 Mar;34(3):328-33. doi: 10.1038/nbt.3471. Epub 2016 Feb 1.
The combination of Cas9, guide RNA and repair template DNA can induce precise gene editing and the correction of genetic diseases in adult mammals. However, clinical implementation of this technology requires safe and effective delivery of all of these components into the nuclei of the target tissue. Here, we combine lipid nanoparticle-mediated delivery of Cas9 mRNA with adeno-associated viruses encoding a sgRNA and a repair template to induce repair of a disease gene in adult animals. We applied our delivery strategy to a mouse model of human hereditary tyrosinemia and show that the treatment generated fumarylacetoacetate hydrolase (Fah)-positive hepatocytes by correcting the causative Fah-splicing mutation. Treatment rescued disease symptoms such as weight loss and liver damage. The efficiency of correction was >6% of hepatocytes after a single application, suggesting potential utility of Cas9-based therapeutic genome editing for a range of diseases.
Cas9、向导RNA和修复模板DNA的组合能够在成年哺乳动物中诱导精确的基因编辑并纠正遗传疾病。然而，这项技术的临床应用需要将所有这些组件安全有效地递送至靶组织的细胞核中。在此，我们将脂质纳米颗粒介导的Cas9 mRNA递送与编码sgRNA和修复模板的腺相关病毒相结合，以诱导成年动物中疾病基因的修复。我们将我们的递送策略应用于人类遗传性酪氨酸血症的小鼠模型，并表明该治疗通过纠正致病性Fah剪接突变产生了富马酰乙酰乙酸水解酶（Fah）阳性的肝细胞。治疗挽救了体重减轻和肝损伤等疾病症状。单次应用后，校正效率超过6%的肝细胞，这表明基于Cas9的治疗性基因组编辑在一系列疾病中具有潜在的应用价值。
