Obstetrics and Gynecology Hospital, State Key Laboratory of Genetic Engineering at School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai, China.
School of Life Sciences, Co-innovation Center of Neuroregeneration, Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong, China.
PLoS Biol. 2020 Mar 30;18(3):e3000686. doi: 10.1371/journal.pbio.3000686. eCollection 2020 Mar.
Compact CRISPR/Cas9 systems that can be packaged into an adeno-associated virus (AAV) hold great promise for gene therapy. Unfortunately, currently available small Cas9 nucleases either display low activity or require a long protospacer adjacent motif (PAM) sequence, limiting their extensive applications. Here, we screened a panel of Cas9 nucleases and identified a small Cas9 ortholog from Staphylococcus auricularis (SauriCas9), which recognizes a simple NNGG PAM, displays high activity for genome editing, and is compact enough to be packaged into an AAV for genome editing. Moreover, the conversion of adenine and cytosine bases can be achieved by fusing SauriCas9 to the cytidine and adenine deaminase. Therefore, SauriCas9 holds great potential for both basic research and clinical applications.
可包装到腺相关病毒 (AAV) 中的紧凑型 CRISPR/Cas9 系统为基因治疗带来了巨大的希望。不幸的是,目前可用的小型 Cas9 核酸酶要么活性低,要么需要长的间隔相邻基序 (PAM) 序列,限制了它们的广泛应用。在这里,我们筛选了一组 Cas9 核酸酶,并从耳葡萄球菌 (SauriCas9) 中鉴定出一种小型 Cas9 同源物,它识别简单的 NNGG PAM,具有高效的基因组编辑活性,并且足够紧凑,可以包装到 AAV 中用于基因组编辑。此外,通过将 SauriCas9 与胞嘧啶和腺嘌呤脱氨酶融合,可以实现腺嘌呤和胞嘧啶碱基的转换。因此,SauriCas9 具有很大的基础研究和临床应用潜力。