SBIR Phase II: Ultra Rapid Genome Engineering in Industrial Yeast Strains

这个小型企业创新研究(SBIR)第二阶段项目的更广泛的影响/商业潜力是能够快速设计酵母来生产燃料、化学品、酶和其他有价值的分子。酵母提供了高价值的生产能力；然而，设计新的菌株是非常复杂和昂贵的-花费大约7500万美元-1亿美元，以目前的工程方法开发需要7-10年。该项目将开发一种酵母基因组工程技术，可以大大减少开发工业生物技术应用的新菌株的成本和时间。这项技术将提高公司使用酵母从可再生原料中高效生产高质量产品的能力，并有助于发展整个工业生物技术市场。这个SBIR第二阶段项目计划发展在工业酵母菌株中进行快速全基因组工程的能力。目前为特定功能(例如，化学或燃料生产)设计电池的技术效率低、成本高且极其耗时。多重自动化基因组工程(MAGE)是一种颠覆性技术，它为以极低的成本设计微生物提供了一个强大的平台。直到最近，MAGE还仅限于一种实验室菌株，具有使MAGE成为可能的关键遗传特征。最近的进展使我们能够将MAGE移植到酵母中，该项目的目标是提高系统的效率，并赋予现有的工业酵母经历MAGE的能力。我们的目标是通过开发一种高通量系统来快速创建和量化菌株变体，从而提高其在酵母中的MAGE效率，从而提高其自身经历MAGE过程的能力。本项目将建立一个系统来鉴定和优化酵母中MAGE所需的遗传特征，以便快速设计出具有特定功能的工业酵母菌株。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase II project is the ability to rapidly engineer yeast for production of fuels, chemicals, enzymes, and other valuable molecules. Yeast offers high value production capabilities; however, engineering new strains is very complex and expensive - costing approximately $75-100 million, and requiring 7-10 years for development with current engineering methods. This project will develop a genome engineering technology for yeast that can substantially reduce the cost and time for developing new strains for industrial biotechnology applications. This technology will increase the ability of companies to use yeast to efficiently produce high-quality products from renewable feedstocks, and help grow the overall industrial biotechnology market. This SBIR Phase II project proposes to develop the ability to perform rapid whole-genome engineering in industrial yeast strains. Current technologies to engineer cells for specific functions (e.g., chemical or fuel production) are inefficient, expensive and extremely time consuming. Multiplex Automated Genome Engineering (MAGE) is a disruptive technology that provides a powerful platform to engineer microorganisms at tremendously reduced cost. Until recently, MAGE was limited to a laboratory strain of E. coli, possessing key genetic features that enable MAGE. Recent advances have allowed us to port MAGE to yeast, and the goal of this project is to increase efficiency of the system and endow existing industrial yeasts with the capacity to undergo MAGE. The goal is to improve MAGE efficiency in yeast by developing a high-throughput system to rapidly create and quantify strain variants for their capacity to undergo the MAGE process itself. This project will establish a system to identify and optimize the requisite genetic features for MAGE in yeast, in order to rapidly engineer industrial yeast strains for specific function.