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在Trichoderma reesei中的基因集群的同时In Vivo组装和有针对性的基因组集成

Yu Fang¹, Xiangfeng Meng¹, Lin Liu¹

¹State Key Laboratory of Microbial Technology, Microbial Technology Institute, Shandong University, No. 72 Binhai Road, Qingdao 266237, P. R. China.

ACS synthetic biology

|February 6, 2025

概括

一种新的方法,即同时在体内组装和多个DNA片段的向基因组集成 (SATIMD),使得Trichoderma reesei.有效的基因工程成为可能. 这种技术促进了大型DNA片段的组装和整合,以增强真菌菌株的发展.

科学领域:

生物技术是生物技术.
合成生物学合成生物学
菌类学菌类学是指菌类学.

背景情况:

特里科德尔马 (Trichoderma reesei) 是细胞质的关键生产者,也是二次代谢物生物合成的潜在宿主.
有效的基因工程方法,特别是用于转移大型DNA片段,对于推进T. reesei的应用至关重要.

研究的目的:

开发一种新的方法,在T. reesei.中同时进行多个DNA片段 (SATIMD) 的in vivo组装和向基因组集成.
证明SATIMD在组装和表达基因以及整合大型基因集群方面的实用性.

主要方法:

开发了SATIMD,将同源重组 (HR) 与CRISPR/Cas9介导的基因组切割相结合.
利用不同的序列重叠长度 (100-500 bp) 进行高效的DNA末端融合.
将组装DNA精确整合到特定的基因组位置.

主要成果:

成功组装和功能表达了T. reesei转录激活器Xyr1用于细胞酶基因.
实现了多达10个DNA片段 (32.7kb) 的单步转化和集成,用于sorbicillinoids生物合成.
验证了HR和CRISPR/Cas9在同时DNA组装和有针对性的集成中的效率.

结论:

SATIMD是T. reesei.中大型异质基因集群组装的强大工具.
这种方法显著促进了真菌菌株的大规模基因工程.
SATIMD增强了T. reesei作为生物技术应用的主机的潜力.

关键词:

克里斯普尔是什么意思?克里斯普尔是什么意思?吃得很飽的吃得很飽的三甲虫reesei 在线阅读同类的重组组合.在活体中,DNA组装.有针对性的基因组整合.

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