Wang Shuai, Ding Yue, Rong Hua, Wang Yu
College of Bioscience and Bioengineering, Jiangxi Agricultural University, Nanchang 330045, China.
Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Nanchang 330045, China.
Curr Issues Mol Biol. 2024 Jan 5;46(1):570-584. doi: 10.3390/cimb46010037.
is a low-GC-content Gram-negative opportunistic pathogen that poses a serious global public health threat. Convenient and rapid genetic manipulation is beneficial for elucidating its pathogenic mechanisms and developing novel therapeutic methods. In this study, we report a new CRISPR-FnCpf1-based two-plasmid system for versatile and precise genome editing in . After identification, this new system prefers to recognize the 5'-TTN-3' (N = A, T, C or G) and the 5'-CTV-3' (V = A, C or G) protospacer-adjacent motif (PAM) sequence and utilize the spacer with lengths ranging from 19 to 25 nt. In direct comparison with the existing CRISPR-Cas9 system, it exhibits approximately four times the targetable range in . Moreover, by employing a tandem dual crRNA expression cassette, the new system can perform large-fragment deletion and simultaneous multiple gene editing, which is difficult to achieve via CRISPR-Cas9. Therefore, the new system is valuable and can greatly expand the genome editing toolbox of .
是一种低GC含量的革兰氏阴性机会致病菌，对全球公共卫生构成严重威胁。便捷快速的基因操作有利于阐明其致病机制并开发新的治疗方法。在本研究中，我们报道了一种基于CRISPR-FnCpf1的新型双质粒系统，用于在[具体物种名称未给出]中进行通用且精确的基因组编辑。经鉴定，该新系统倾向于识别5'-TTN-3'（N = A、T、C或G）和5'-CTV-3'（V = A、C或G）的原间隔序列临近基序（PAM）序列，并利用长度为19至25 nt的间隔序列。与现有的CRISPR-Cas9系统直接比较，它在[具体物种名称未给出]中的靶向范围约为四倍。此外，通过采用串联双crRNA表达盒，新系统可进行大片段缺失和同时多个基因编辑，这是CRISPR-Cas9难以实现的。因此，新系统具有重要价值，可极大地扩展[具体物种名称未给出]的基因组编辑工具箱。
