Frontiers Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin, China.
Haihe Laboratory of Sustainable Chemical Transformations, Tianjin, China.
Nat Commun. 2024 Nov 13;15(1):9833. doi: 10.1038/s41467-024-54196-z.
The multifunctional proteins of class 2 CRISPR systems such as Cas9, have been employed to activate cryptic biosynthetic gene clusters (BGCs) in Streptomyces, which represent a large and hidden reservoir of natural products. However, such approaches are not applicable to most Streptomyces strains with reasons to be comprehended. Inspired by the prevalence of the class 1 subtype especially the type I-E CRISPR system in Streptomyces, here we report the development of the type I-E CRISPR system into a series of transcriptional regulation tools. We further demonstrate the effectiveness of such activators in nine phylogenetically distant Streptomyces strains. Using these tools, we successfully activate 13 out of 21 BGCs and lead to the identification and characterization of one polyketide, one Ripp and three alkaloid products. Our work is expected to have a profound impact and to facilitate the discovery of numerous structurally diverse compounds from Streptomyces.
2 类 CRISPR 系统的多功能蛋白，如 Cas9，已被用于激活链霉菌中隐蔽的生物合成基因簇（BGCs），这些基因簇代表了一个庞大而隐藏的天然产物资源库。然而，由于某些原因，这些方法并不适用于大多数链霉菌菌株。受 1 类亚型特别是 I-E 型 CRISPR 系统在链霉菌中普遍存在的启发，我们在这里报告将 I-E 型 CRISPR 系统开发成一系列转录调控工具。我们进一步证明了这些激活子在九株系统发育上相距较远的链霉菌菌株中的有效性。使用这些工具，我们成功激活了 21 个 BGC 中的 13 个，并鉴定和表征了一个聚酮、一个 Ripp 和三个生物碱产物。我们的工作预计将产生深远的影响，并促进从链霉菌中发现众多结构多样的化合物。
