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A supernumerary designer chromosome for modular in vivo pathway assembly in Saccharomyces cerevisiae
Nucleic Acids Research ( IF 14.9 ) Pub Date : 2021-01-11 , DOI: 10.1093/nar/gkaa1167
Eline D Postma 1 , Sofia Dashko 1 , Lars van Breemen 1 , Shannara K Taylor Parkins 1 , Marcel van den Broek 1 , Jean-Marc Daran 1 , Pascale Daran-Lapujade 1
Affiliation  

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.

中文翻译:

用于酿酒酵母中模块化体内途径组装的多余设计染色体。

为了可持续地生产化学药品和药品,建造微生物细胞工厂需要大量的基因组工程。使用酿酒酵母,这项研究提出了合成新染色体作为正交表达平台,用于重新连接天然细胞过程和实现新功能。利用酿酒酵母强大的同源重组能力,可以在单个转化中从多达44个转录单位大小的片段重新组装50和100 kb的模块化新染色体。这些集会非常高效并且忠实于他们的计算机技术设计。由非编码DNA制成的新染色体可以稳定地复制和分离,而不管其大小如何,而不会影响宿主的生理。这些非编码新染色体已成功用作必需糖酵解途径的着陆垫和独家表达平台。这项工作推DNA的装配在极限酿酒酵母和铺平了道路从头设计的染色体为模块化基因组工程平台酿酒酵母
更新日期:2021-02-22
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