Gao Jie, Wang Hengyi, Sun Jingtao, Tang Hongjie, Yang Yuhan, Li Qi
College of Life Sciences, Sichuan Normal University, Chengdu, 610101, China.
Synth Syst Biotechnol. 2025 Jul 2;10(4):1215-1223. doi: 10.1016/j.synbio.2025.06.012. eCollection 2025 Dec.
Existing CRISPR-based genome editing techniques for () are limited due to the large size of the gene. IscB, a recently reported DNA nuclease, is one-third the size of Cas9, making it a potential tool for genome editing; however, its application in remains unexplored. In this study, two IscB and enhanced IscB (enIscB)-based genome editing systems, named pBsuIscB and pBsuenIscB were established in SCK6, and their deletion efficiencies ranging from 13.3 % to 100 %. Compared to the pBsuIscB system, the pBsuenIscB system showed higher deletion efficiency, inducing the deletion of a large genomic fragment with a single ωRNA. Additionally, the pBsuenIscB system could integrate both single-copy and multi-copy genes in the SCK6 genome. Lastly, the pBsuenIscB system could efficiently conduct a second round of genome editing in SCK6. This study indicates that IscB can be used for genome editing in , enabling the efficient construction of engineered strains for large-scale biomolecule production.
由于（）基因的尺寸较大，现有的基于CRISPR的基因组编辑技术受到限制。IscB是最近报道的一种DNA核酸酶，其大小仅为Cas9的三分之一，使其成为基因组编辑的潜在工具；然而，其在（）中的应用尚未得到探索。在本研究中，在（）SCK6中建立了两个基于IscB和增强型IscB（enIscB）的基因组编辑系统，分别命名为pBsuIscB和pBsuenIscB，它们的缺失效率在13.3%至100%之间。与pBsuIscB系统相比，pBsuenIscB系统显示出更高的缺失效率，能够通过单个ωRNA诱导大片段基因组的缺失。此外，pBsuenIscB系统能够在（）SCK6基因组中整合单拷贝和多拷贝（）基因。最后，pBsuenIscB系统能够在（）SCK6中高效地进行第二轮基因组编辑。本研究表明，IscB可用于（）的基因组编辑，从而能够高效构建用于大规模生物分子生产的工程（）菌株。
