Hren Andrew, Alexander William G, Abraham Joshua P, Tumen-Velasquez Melissa P, Melesse Vergara Michael, Guss Adam M, Pfleger Brian F, Fox Jerome M, Eckert Carrie A
Department of Chemical and Biological Engineering, University of Colorado, Boulder, Colorado 80304, United States.
Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831, United States.
ACS Synth Biol. 2025 Aug 15;14(8):3258-3264. doi: 10.1021/acssynbio.5c00370. Epub 2025 Aug 4.
Cyanobacteria are promising microbial platforms for a diverse set of biotechnology applications, from living materials to photosynthetic chemical production, but are less well characterized than commonly engineered microbes such as . This study facilitates genetic engineering in sp. PCC 7002, a fast-growing, halotolerant, and naturally competent strain, by identifying ten native methylation motifs and designing shuttle strains that mimic the native methylation state by expressing a subset of heterologous methyltransferases. DNA methylation in with as few as two active methyltransferases increased transformation efficiency up to 30-fold across four distinct integration sites in PCC 7002. This work provides an experimental framework to bypass native restriction-modification systems for efficient genome editing and metabolic engineering in nonmodel bacteria.
蓝藻细菌是用于从生物材料到光合化学品生产等多种生物技术应用的有前景的微生物平台，但与诸如[此处原文缺失常见工程微生物名称]等常见工程微生物相比，其特征描述较少。本研究通过鉴定十个天然甲基化基序并设计穿梭菌株，促进了集胞藻属PCC 7002（一种生长迅速、耐盐且天然感受态的菌株）中的基因工程，所述穿梭菌株通过表达异源甲基转移酶的一个子集来模拟天然甲基化状态。在集胞藻属中，仅用两种活性甲基转移酶进行DNA甲基化，就在PCC 7002的四个不同整合位点将转化效率提高了30倍。这项工作提供了一个实验框架，用于绕过天然限制修饰系统，以在非模式细菌中进行高效的基因组编辑和代谢工程。
