School of Physical Science and Technology, ShanghaiTech University, Shanghai 201210, China.
University of Chinese Academy of Sciences, Beijing 100049, China.
ACS Synth Biol. 2022 Sep 16;11(9):3049-3057. doi: 10.1021/acssynbio.2c00248. Epub 2022 Aug 24.
is a major human pathogen that causes a variety of infections, including life-threatening diseases. Research on is constrained by complex and limited genetic manipulation methods. Here, we report a CRISPR/Cpf1-mediated system, pCpfSA, for rapid and versatile genome editing in . In direct comparison with the existing CRISPR/Cas9-mediated genome-editing system, the pCpfSA system exhibits enhanced colony-forming units (CFUs) after editing and an expanded targetable range with comparable editing efficiency. Given the precursor crRNA (pre-crRNA) processing activity of Cpf1, the pCpfSA system also allows multiplex gene editing and large-fragment DNA knockout simply by introducing two crRNAs and the corresponding donor templates, which is difficult to achieve using the CRISPR/Cas9 system, thereby greatly expanding the genome editor toolbox for .
是一种主要的人类病原体，可引起多种感染，包括危及生命的疾病。对 的研究受到复杂和有限的遗传操作方法的限制。在这里，我们报告了一种 CRISPR/Cpf1 介导的系统 pCpfSA，用于 在 中快速和多功能的基因组编辑。与现有的 CRISPR/Cas9 介导的基因组编辑系统直接比较，pCpfSA 系统在编辑后表现出增强的集落形成单位 (CFUs)，并且具有可比的编辑效率的可靶向范围扩大。鉴于 Cpf1 的前体 crRNA (pre-crRNA) 加工活性，pCpfSA 系统还允许通过引入两个 crRNA 和相应的供体模板进行多重基因编辑和大片段 DNA 敲除，这使用 CRISPR/Cas9 系统很难实现，从而大大扩展了 基因组编辑工具包。
