Wang Xin, Tang Yu, Lu Jian, Shao Yanjiao, Qin Xuan, Li Yongmei, Wang Liren, Li Dali, Liu Mingyao
Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China.
Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China.
Biochem Pharmacol. 2016 Apr 1;105:80-90. doi: 10.1016/j.bcp.2016.03.001. Epub 2016 Mar 3.
A bacterial CRISPR-associated protein-9 nuclease (CRISPR/Cas9) from Streptococcus pyogenes has generated considerable excitement as a new tool to edit the targeted genome. Cytochrome P450 (CYP) 2E1 not only plays an important role in the xenobiotic metabolism and chemical toxicity, but also is involved in many kinds of diseases, such as alcoholic liver diseases and diabetes. Despite its importance, few animal models are used to predict CYP2E1 properties in physiology, pathology, as well as carcinogen activation. To establish a novel model for investigating the functions of CYP2E1 in vivo, this study has successfully generated the Cyp2e1 knockout (KO) rat model without detectable off-target effects using CRISPR/Cas9 system. The Cyp2e1 KO rats were viable and fertile and did not display any obvious physiological abnormities. The absent expression of CYP2E1 in KO rats also resulted in inactive behaviors in the metabolism of CYP2E1 substrates. The Cyp2e1 KO rats as a novel and available rodent animal model provide a powerful tool for the study of CYP2E1 in the chemical metabolism, toxicity, carcinogenicity, and its core factor in drug-drug interactions.
来自化脓性链球菌的一种细菌CRISPR相关蛋白9核酸酶（CRISPR/Cas9）作为一种编辑靶向基因组的新工具，引发了广泛关注。细胞色素P450（CYP）2E1不仅在异源物质代谢和化学毒性中发挥重要作用，还参与多种疾病，如酒精性肝病和糖尿病。尽管其很重要，但很少有动物模型用于预测CYP2E1在生理、病理以及致癌物激活方面的特性。为建立一种用于在体内研究CYP2E1功能的新型模型，本研究利用CRISPR/Cas9系统成功构建了无明显脱靶效应的Cyp2e1基因敲除（KO）大鼠模型。Cyp2e1 KO大鼠存活且可育，未表现出任何明显的生理异常。KO大鼠中CYP2E1的缺失表达也导致了CYP2E1底物代谢的无活性行为。Cyp2e1 KO大鼠作为一种新型且可用的啮齿动物模型，为研究CYP2E1在化学代谢、毒性、致癌性及其在药物相互作用中的核心因素提供了有力工具。
