Sanofi-Aventis Deutschland GmbH, Global CMC Microbial Platform, USP Development - Molecular Biology, Frankfurt am Main, Germany.
Microb Biotechnol. 2023 May;16(5):1000-1010. doi: 10.1111/1751-7915.14234. Epub 2023 Mar 16.
A broad variety of biomolecules is industrially produced in bacteria and yeasts. These microbial expression hosts can be optimized through genetic engineering using CRISPR tools. Here, we designed and characterized such a modular genome editing system based on the Cas12a-like RNA-guided nuclease MAD7 in Escherichia coli. This system enables the efficient generation of single nucleotide polymorphisms (SNPs) or gene deletions and can directly be used with donor DNA from benchtop DNA assembly to increase throughput. We combined multiple edits to engineer an E. coli strain with reduced overflow metabolism and increased plasmid yield, highlighting the versatility and industrial applicability of this approach.
多种多样的生物分子在细菌和酵母中被工业化生产。这些微生物表达宿主可以通过使用 CRISPR 工具的基因工程进行优化。在这里，我们设计并表征了一种基于大肠杆菌 Cas12a 样 RNA 指导的核酸酶 MAD7 的模块化基因组编辑系统。该系统能够高效地产生单核苷酸多态性（SNP）或基因缺失，并且可以直接与来自台式 DNA 组装的供体 DNA 一起使用，以提高通量。我们结合了多个编辑来工程改造大肠杆菌菌株，以减少溢出代谢并增加质粒产量，突出了这种方法的多功能性和工业适用性。
