MAD7 is an engineered class 2 type V-A CRISPR-Cas (Cas12a/Cpf1) system isolated from Eubacterium rectale. Analogous to Cas9, it is an RNA-guided nuclease with demonstrated gene editing activity in Escherichia coli and yeast cells. Here, we report that MAD7 is capable of generating indels and fluorescent gene tagging of endogenous genes in human HCT116 and U2OS cancer cell lines, respectively. In addition, MAD7 is highly proficient in generating indels, small DNA insertions (23 bases), and larger integrations ranging from 1 to 14 kb in size in mouse and rat embryos, resulting in live-born transgenic animals. Due to the different protospacer adjacent motif requirement, small-guide RNA, and highly efficient targeted gene disruption and insertions, MAD7 can expand the CRISPR toolbox for genome enginnering across different systems and model organisms.

中文翻译：

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用于人类细胞、小鼠和大鼠模型生成的 ErCas12a CRISPR-MAD7。


MAD7 是从直肠真杆菌中分离出来的工程 2 类 VA CRISPR-Cas (Cas12a/Cpf1) 系统。与 Cas9 类似，它是一种 RNA 引导的核酸酶，在大肠杆菌和酵母细胞中具有基因编辑活性。在这里，我们报告 MAD7 能够分别在人 HCT116 和 U2OS 癌细胞系中生成内源基因的插入缺失和荧光基因标签。此外，MAD7 非常擅长在小鼠和大鼠胚胎中生成插入缺失、小 DNA 插入（23 个碱基）以及大小从 1 到 14 kb 的较大整合，从而产生活生的转基因动物。由于不同的原型间隔子相邻基序要求、小引导 RNA 以及高效的靶向基因破坏和插入，MAD7 可以扩展 CRISPR 工具箱，用于跨不同系统和模型生物的基因组工程。