通过CRISPR-Cas9在酵母中多重整合协同等位基因和代谢途径的方法
Method for Multiplexed Integration of Synergistic Alleles and Metabolic Pathways in Yeasts via CRISPR-Cas9.
作者信息
Walter Jessica M, Schubert Max G, Kung Stephanie H, Hawkins Kristy, Platt Darren M, Hernday Aaron D, Mahatdejkul-Meadows Tina, Szeto Wayne, Chandran Sunil S, Newman Jack D, Horwitz Andrew A
机构信息
Amyris, Inc., Emeryville, CA, USA.
出版信息
Methods Mol Biol. 2019;2049:39-72. doi: 10.1007/978-1-4939-9736-7_3.
CRISPR-Cas has proven to be a powerful tool for precision genetic engineering in a variety of difficult genetic systems. In the highly tractable yeast S. cerevisiae, CRISPR-Cas can be used to conduct multiple engineering steps in parallel, allowing for engineering of complex metabolic pathways at multiple genomic loci in as little as 1 week. In addition, CRISPR-Cas can be used to consolidate multiple causal alleles into a single strain, bypassing the laborious traditional methods using marked constructs, or mating. These tools compress the engineering timeline sixfold or more, greatly increasing the productivity of the strain engineer.
CRISPR-Cas已被证明是一种在各种复杂遗传系统中进行精确基因工程的强大工具。在易于操作的酿酒酵母中，CRISPR-Cas可用于并行进行多个工程步骤，从而能够在短短1周内对多个基因组位点的复杂代谢途径进行工程改造。此外，CRISPR-Cas可用于将多个致病等位基因整合到单个菌株中，绕过使用标记构建体或交配的繁琐传统方法。这些工具将工程时间缩短了六倍或更多，极大地提高了菌株工程师的工作效率。
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