A Highly Characterized Yeast Toolkit for Modular, Multipart Assembly
A Highly Characterized Yeast Toolkit for Modular, Multipart Assembly
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DOI：
10.1021/sb500366v
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发表时间：
2015-09-01
影响因子：
4.7
通讯作者：
Dueber, John E.
中科院分区：
文献类型：
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作者：
Lee, Michael E.;DeLoache, William C.;Dueber, John E.
酿酒酵母由于其在工业发酵中的悠久历史而成为合成生物学越来越有吸引力的宿主。然而，直到最近，大多数合成生物学系统都集中在细菌上。虽然有大量关于酵母生物学的资源和文献，但导航和提取工程应用所需的工具可能令人生畏。在这里，我们提出了一个多功能的工程平台，酵母，其中包含一个快速，模块化的组装方法和一套基本的特征部分。该平台提供了一个框架，在其中创建新的设计，以及启动子，终止子，降解标签和拷贝数的数据，以告知这些设计。此外，我们描述了用于直接对酵母染色体进行修饰的基因组编辑工具，由于表达的可变性降低，我们发现这种工具比质粒更好。有了这个工具包，我们努力简化工程酵母的过程，通过标准化的物理操作和建议的最佳实践，共同将使材料和数据从一个组到另一个组的更直接的翻译。此外，通过减轻研究人员对技术细节的负担，他们可以专注于实验设计的更高层次。
Saccharomyces cerevisiae is an increasingly attractive host for synthetic biology because of its long history in industrial fermentations. However, until recently, most synthetic biology systems have focused on bacteria. While there is a wealth of resources and literature about the biology of yeast, it can be daunting to navigate and extract the tools needed for engineering applications. Here we present a versatile engineering platform for yeast, which contains both a rapid, modular assembly method and a basic set of characterized parts. This platform provides a framework in which to create new designs, as well as data on promoters, terminators, degradation tags, and copy number to inform those designs. Additionally, we describe genome-editing tools for making modifications directly to the yeast chromosomes, which we find preferable to plasmids due to reduced variability in expression. With this toolkit, we strive to simplify the process of engineering yeast by standardizing the physical manipulations and suggesting best practices that together will enable more straightforward translation of materials and data from one group to another. Additionally, by relieving researchers of the burden of technical details, they can focus on higher-level aspects of experimental design.