CRISPR/Cas 介导的链霉菌基因组编辑。
CRISPR/Cas-Mediated Genome Editing of Streptomyces.
机构信息
Molecular Engineering Lab, Institute of Molecular and Cell Biology, A*STAR, Singapore, Singapore.
Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan, Miaoli, Taiwan.
出版信息
Methods Mol Biol. 2022;2479:207-225. doi: 10.1007/978-1-0716-2233-9_14.
Streptomyces are an important source and reservoir of natural products with diverse applications in medicine, agriculture, and food. Engineered Streptomyces strains have also proven to be functional chassis for the discovery and production of bioactive compounds and enzymes. However, genetic engineering of Streptomyces is often laborious and time-consuming. Here we describe protocols for CRISPR/Cas-mediated genome editing of Streptomyces. Starting from the design and assembly of all-in-one CRISPR/Cas constructs for efficient double-strand break-mediated genome editing, we also present protocols for intergeneric conjugation, CRISPR/Cas plasmid curing, and validation of edited strains.
链霉菌是天然产物的重要来源和储存库，在医学、农业和食品领域有广泛的应用。经过基因工程改造的链霉菌菌株也被证明是发现和生产生物活性化合物和酶的有效底盘。然而，链霉菌的遗传工程通常既费力又耗时。在这里，我们描述了用于链霉菌的 CRISPR/Cas 介导的基因组编辑的方案。从用于高效双链断裂介导的基因组编辑的一体式 CRISPR/Cas 构建体的设计和组装开始，我们还介绍了用于种间接合、CRISPR/Cas 质粒消除和编辑菌株验证的方案。
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