Improving the Transformation Efficiency of Synechococcus sp. PCC 7002 via Methylome-Guided Premethylation of DNA

IF 3.9 2区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

ACS Synthetic Biology Pub Date : 2025-08-04 DOI:10.1021/acssynbio.5c00370

Andrew Hren, William G. Alexander, Joshua P. Abraham, Melissa P. Tumen-Velasquez, Michael Melesse Vergara, Adam M. Guss, Brian F. Pfleger*, Jerome M. Fox* and Carrie A. Eckert*,

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引用次数: 0

Abstract

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 Escherichia coli. This study facilitates genetic engineering in Synechococcus 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 E. coli 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.

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甲基组引导DNA预甲基化提高聚球菌pcc7002转化效率

蓝藻是一种很有前途的微生物平台，可用于多种生物技术应用，从生物材料到光合化学生产，但与大肠杆菌等普通工程微生物相比，蓝藻的特征不那么明显。本研究通过鉴定10个天然甲基化基元，并设计通过表达一组异源甲基转移酶来模拟天然甲基化状态的穿梭菌株，促进了聚球菌PCC 7002的基因工程，PCC 7002是一种快速生长、耐盐、自然胜任的菌株。在大肠杆菌中，只要有两种活性甲基转移酶，就能将pcc7002中四个不同整合位点的DNA甲基化效率提高30倍。这项工作提供了一个实验框架，可以绕过天然限制性修饰系统，在非模式细菌中进行高效的基因组编辑和代谢工程。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

ACS Synthetic Biology 生物-

CiteScore

8.00

自引率

10.60%

发文量

380

审稿时长

6-12 weeks

期刊介绍： The journal is particularly interested in studies on the design and synthesis of new genetic circuits and gene products; computational methods in the design of systems; and integrative applied approaches to understanding disease and metabolism. Topics may include, but are not limited to: Design and optimization of genetic systems Genetic circuit design and their principles for their organization into programs Computational methods to aid the design of genetic systems Experimental methods to quantify genetic parts, circuits, and metabolic fluxes Genetic parts libraries: their creation, analysis, and ontological representation Protein engineering including computational design Metabolic engineering and cellular manufacturing, including biomass conversion Natural product access, engineering, and production Creative and innovative applications of cellular programming Medical applications, tissue engineering, and the programming of therapeutic cells Minimal cell design and construction Genomics and genome replacement strategies Viral engineering Automated and robotic assembly platforms for synthetic biology DNA synthesis methodologies Metagenomics and synthetic metagenomic analysis Bioinformatics applied to gene discovery, chemoinformatics, and pathway construction Gene optimization Methods for genome-scale measurements of transcription and metabolomics Systems biology and methods to integrate multiple data sources in vitro and cell-free synthetic biology and molecular programming Nucleic acid engineering.