Chen Siyu, Xie Wanhua, Liu Zhiquan, Shan Huanhuan, Chen Mao, Song Yuning, Yu Hao, Lai Liangxue, Li Zhanjun
Key Laboratory of Zoonosis Research, Ministry of Education, College of Animal Science, Jilin University, Changchun 130062, China.
The Precise Medicine Center, Shenyang Medical College, Shenyang, China.
Mol Ther Nucleic Acids. 2020 Sep 4;21:1062-1073. doi: 10.1016/j.omtn.2020.07.037. Epub 2020 Jul 31.
CRISPR-Cas9-mediated gene knockout and base-editing-associated induction of STOP codons (iSTOP) have been widely used to exterminate the function of a coding gene, while they have been reported to exhibit side effects. In this study, we propose a novel and practical alternative method referred to as CRISPR Start-Loss (CRISPR-SL), which eliminates gene expression by utilizing both adenine base editors (ABEs) and cytidine base editors (CBEs) to disrupt the initiation codon (ATG). CRISPR-SL has been verified to be a feasible strategy on the cellular and embryonic levels (mean editing efficiencies up to 30.67% and 73.50%, respectively) and in two rabbit models mimicking Otc deficiency (Otc gene) and long hair economic traits (Fgf5 gene).
CRISPR-Cas9介导的基因敲除以及与碱基编辑相关的终止密码子诱导（iSTOP）已被广泛用于消除编码基因的功能，然而据报道它们存在副作用。在本研究中，我们提出了一种新颖且实用的替代方法，称为CRISPR起始密码子缺失（CRISPR-SL），该方法通过利用腺嘌呤碱基编辑器（ABE）和胞嘧啶碱基编辑器（CBE）来破坏起始密码子（ATG）从而消除基因表达。CRISPR-SL已在细胞和胚胎水平（平均编辑效率分别高达30.67%和73.50%）以及两种模拟鸟氨酸转氨甲酰酶缺乏症（Otc基因）和长毛经济性状（Fgf5基因）的兔模型中被验证为一种可行的策略。
