Hubei International Scientific and Technological Cooperation Base of Traditional Fermented Foods, Huazhong Agricultural University, Wuhan, Hubei Province 430070, China.
College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei Province 430070, China.
ACS Synth Biol. 2023 Jul 21;12(7):2147-2156. doi: 10.1021/acssynbio.3c00229. Epub 2023 Jul 10.
Random base editing is regarded as a fundamental method for accelerating the genomic evolution in both scientific research and industrial applications. In this study, we designed a modular interaction-based dual base editor (MIDBE) that assembled a DNA helicase and various base editors through dockerin/cohesin-mediated protein-protein interactions, resulting in a self-assembled MIDBE complex capable of editing bases at any locus in the genome. The base editing type of MIDBE can be readily controlled by the induction of cytidine or/and adenine deaminase gene expression. MIDBE exhibited the highest editing efficiency 2.3 × 10 times greater than the native genomic mutation rate. To evaluate the potential of MIDBE in genomic evolution, we developed a removable plasmid-based MIDBE tool, which led to a remarkable 977.1% increase of lovastatin production in HJ11. MIDBE represents the first biological tool for generating and accumulating base mutations in Monascus chromosome and also offers a bottom-up strategy for designing the base editor.
随机碱基编辑被认为是加速科学研究和工业应用中基因组进化的一种基本方法。在这项研究中，我们设计了一种基于模块化相互作用的双碱基编辑器（MIDBE），它通过 dockerin/cohesin 介导的蛋白质-蛋白质相互作用将 DNA 解旋酶和各种碱基编辑器组装在一起，形成一个能够在基因组中的任何位点编辑碱基的自组装 MIDBE 复合物。MIDBE 的碱基编辑类型可以通过诱导胞嘧啶或/和腺嘌呤脱氨酶基因表达来轻松控制。MIDBE 表现出的编辑效率比天然基因组突变率高 2.3×10 倍。为了评估 MIDBE 在基因组进化中的潜力，我们开发了一种可移动的基于质粒的 MIDBE 工具，这导致 HJ11 中洛伐他汀产量显著增加了 977.1%。MIDBE 代表了在红曲菌染色体中产生和积累碱基突变的第一个生物工具，也为设计碱基编辑器提供了一种自下而上的策略。
