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在Coleophoma empetri中由CRISPR/Cas9介导的可视化和无标记物基因编辑平台

Li Xie^1,2,3, Ping Men^1,2,3, Yu Zhou^1,2,3,4

¹State Key Laboratory of Photoelectric Conversion and Utilization of Solar Energy, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, 266101, China.

Applied microbiology and biotechnology

|February 25, 2026

概括

为Coleophoma empetri开发了一个新的可可视化,无标记物基因编辑平台,使FR901379的生产能够得到增强. 这个系统加速了工业真菌菌株的代谢工程和功能基因组学.

科学领域:

菌类学菌类学是指菌类学.
分子生物学分子生物学
生物技术是生物技术.

背景情况:

米卡金是一种赤甘丁抗真菌药物,是 FR901379 的衍生物,由 Coleophoma empetri 生产.
工业C. empetri菌株的基因操纵是具有挑战性的,因为有限的中性位点和可选择的标记物.

研究的目的:

开发一个可视化和无标记基因编辑平台,用于工业C. empetri菌株.
为了促进C. empetri. 的代谢工程和功能基因组学.

主要方法:

在C. empetri的黑色素生物合成基因集群 (cemelA,cemelR) 的表征 MEFC09.09.
使用 luciferase reporter 设计和验证两个中性基因组位点.
开发一种CRISPR/Cas9介导的无标记物基因编辑系统.
优化原生质度以实现高效的转化.

主要成果:

在cemelA和cemelR位点确定了两个高表达的中性位点.
在没有抗生素查的情况下,CRISPR/Cas9系统实现了12.4%的转化率.
转录激活剂McfJ在中性部位的过度表达增加了FR901379标位的3.3倍 (1807.8毫克/升).

结论:

已经建立了一个新的,高效的基因编辑平台,用于C. empetri.
该平台通过向基因表达显著增强FR901379的产生.
该系统适用于推进工业丝状真菌中的代谢工程和功能基因组学.

关键词:

这就是CRISPR/Cas9的作用.工业丝状真菌工业丝状真菌没有标记物的查.在米卡菌中,有菌素.可视化的中立地点.

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