清脆的
基因组编辑
计算生物学
同色链霉菌
多路复用
多路复用
生物
计算机科学
胞苷脱氨酶
Cas9
基因组
链霉菌
基因
遗传学
电信
细菌
作者
Christopher M. Whitford,Tetiana Gren,Emilia Palazzotto,Sang Yup Lee,Yaojun Tong,Tilmann Weber
标识
DOI:10.1021/acssynbio.3c00188
摘要
CRISPR tools, especially Cas9n-sgRNA guided cytidine deaminase base editors such as CRISPR-BEST, have dramatically simplified genetic manipulation of streptomycetes. One major advantage of CRISPR base editing technology is the possibility to multiplex experiments in genomically instable species. Here, we demonstrate scaled up Csy4 based multiplexed genome editing using CRISPR-mcBEST in Streptomyces coelicolor. We evaluated the system by simultaneously targeting 9, 18, and finally all 28 predicted specialized metabolite biosynthetic gene clusters in a single experiment. We present important insights into the performance of Csy4 based multiplexed genome editing at different scales. Using multiomics analysis, we investigated the systems wide effects of such extensive editing experiments and revealed great potentials and important bottlenecks of CRISPR-mcBEST. The presented analysis provides crucial data and insights toward the development of multiplexed base editing as a novel paradigm for high throughput engineering of Streptomyces chassis and beyond.
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