Riley Lauren A, Guss Adam M
Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN, 37831, USA.
Bredesen Center, University of Tennessee, Knoxville, TN, 37996, USA.
Biotechnol Biofuels. 2021 Jan 25;14(1):30. doi: 10.1186/s13068-020-01872-z.
Non-model microorganisms often possess complex phenotypes that could be important for the future of biofuel and chemical production. They have received significant interest the last several years, but advancement is still slow due to the lack of a robust genetic toolbox in most organisms. Typically, "domestication" of a new non-model microorganism has been done on an ad hoc basis, and historically, it can take years to develop transformation and basic genetic tools. Here, we review the barriers and solutions to rapid development of genetic transformation tools in new hosts, with a major focus on Restriction-Modification systems, which are a well-known and significant barrier to efficient transformation. We further explore the tools and approaches used for efficient gene deletion, DNA insertion, and heterologous gene expression. Finally, more advanced and high-throughput tools are now being developed in diverse non-model microbes, paving the way for rapid and multiplexed genome engineering for biotechnology.
非模式微生物通常具有复杂的表型，这对生物燃料和化学品生产的未来可能至关重要。在过去几年中，它们受到了广泛关注，但由于大多数生物体缺乏强大的遗传工具箱，进展仍然缓慢。通常，新的非模式微生物的“驯化”是临时进行的，从历史上看，开发转化和基本遗传工具可能需要数年时间。在这里，我们回顾了新宿主中遗传转化工具快速开发的障碍和解决方案，主要关注限制修饰系统，这是高效转化的一个众所周知且重要的障碍。我们进一步探索了用于高效基因缺失、DNA插入和异源基因表达的工具和方法。最后，现在正在多种非模式微生物中开发更先进和高通量的工具，为生物技术的快速和多重基因组工程铺平了道路。
