Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA.
Gladstone Institutes, San Francisco, CA 94158, USA.
Science. 2018 Oct 12;362(6411):236-239. doi: 10.1126/science.aau5138. Epub 2018 Sep 6.
Cas12a (Cpf1) is a CRISPR-associated nuclease with broad utility for synthetic genome engineering, agricultural genomics, and biomedical applications. Although bacteria harboring CRISPR-Cas9 or CRISPR-Cas3 adaptive immune systems sometimes acquire mobile genetic elements encoding anti-CRISPR proteins that inhibit Cas9, Cas3, or the DNA-binding Cascade complex, no such inhibitors have been found for CRISPR-Cas12a. Here we use a comprehensive bioinformatic and experimental screening approach to identify three different inhibitors that block or diminish CRISPR-Cas12a-mediated genome editing in human cells. We also find a widespread connection between CRISPR self-targeting and inhibitor prevalence in prokaryotic genomes, suggesting a straightforward path to the discovery of many more anti-CRISPRs from the microbial world.
Cas12a（Cpf1）是一种 CRISPR 相关的核酸酶，在合成基因组工程、农业基因组学和生物医学应用方面具有广泛的用途。尽管携带 CRISPR-Cas9 或 CRISPR-Cas3 适应性免疫系统的细菌有时会获得编码抗 Cas9、Cas3 或 DNA 结合级联复合物的移动遗传元件，但尚未发现针对 CRISPR-Cas12a 的此类抑制剂。在这里，我们使用全面的生物信息学和实验筛选方法来鉴定三种不同的抑制剂，它们可以阻断或削弱 CRISPR-Cas12a 在人类细胞中的基因组编辑。我们还发现 CRISPR 自我靶向与原核基因组中抑制剂流行之间存在广泛的联系，这表明从微生物世界中发现更多抗 CRISPR 的方法非常直接。
