Although only 6 years old, the CRISPR system has blossomed into a tool for rapid, on-demand genome engineering and gene regulation in Saccharomyces cerevisiae. In this minireview, we discuss fundamental CRISPR technologies, tools to improve the efficiency and capabilities of gene targeting, and cutting-edge techniques to explore gene editing and transcriptional regulation at genome scale using pooled approaches. The focus is on applications to metabolic engineering with topics including development of techniques to edit the genome in multiplex, tools to enable large numbers of genetic modifications using pooled single-guide RNA libraries and efforts to enable programmable transcriptional regulation using endonuclease-null Cas enzymes.

中文翻译：

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在酿酒酵母中使用 CRISPR/Cas 增强基因组工程和调控的规模和范围。


尽管只有 6 年的历史，CRISPR 系统已发展成为一种在酿酒酵母中进行快速、按需基因组工程和基因调控的工具。在这篇小综述中，我们讨论了基本的 CRISPR 技术、提高基因打靶效率和能力的工具，以及使用汇总方法探索基因组规模的基因编辑和转录调控的尖端技术。重点是代谢工程的应用，主题包括开发多重基因组编辑技术、使用合并的单引导 RNA 文库实现大量遗传修饰的工具，以及使用核酸内切酶无效 Cas 酶实现可编程转录调控的努力。