Department of Biotechnology, Delft University of Technology, van der Maasweg 9, 2627HZ Delft, The Netherlands.
Nucleic Acids Res. 2021 Feb 22;49(3):1769-1783. doi: 10.1093/nar/gkaa1167.
The construction of microbial cell factories for sustainable production of chemicals and pharmaceuticals requires extensive genome engineering. Using Saccharomyces cerevisiae, this study proposes synthetic neochromosomes as orthogonal expression platforms for rewiring native cellular processes and implementing new functionalities. Capitalizing the powerful homologous recombination capability of S. cerevisiae, modular neochromosomes of 50 and 100 kb were fully assembled de novo from up to 44 transcriptional-unit-sized fragments in a single transformation. These assemblies were remarkably efficient and faithful to their in silico design. Neochromosomes made of non-coding DNA were stably replicated and segregated irrespective of their size without affecting the physiology of their host. These non-coding neochromosomes were successfully used as landing pad and as exclusive expression platform for the essential glycolytic pathway. This work pushes the limit of DNA assembly in S. cerevisiae and paves the way for de novo designer chromosomes as modular genome engineering platforms in S. cerevisiae.
为了可持续地生产化学品和药品，需要构建微生物细胞工厂，这需要广泛的基因组工程。本研究利用酿酒酵母提出了合成新染色体，作为重新布线天然细胞过程和实现新功能的正交表达平台。利用酿酒酵母强大的同源重组能力，我们可以在一次转化中从头组装多达 44 个转录单元大小的片段，构建 50 和 100 kb 的模块化新染色体。这些组装非常高效，并且与它们的计算机设计高度一致。由非编码 DNA 组成的新染色体可以稳定复制和分离，无论其大小如何，而不会影响宿主的生理机能。这些非编码新染色体可成功用作着陆垫和必需糖酵解途径的专用表达平台。这项工作推动了酿酒酵母中 DNA 组装的极限，并为从头设计的染色体作为酿酒酵母中的模块化基因组工程平台铺平了道路。
Zotero 插件