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Updated: Sep 17, 2025

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在Paecilomyces variotii中开发CRISPR/Cas9 RNP介导的基因工程系统

Hui-Gang Han¹, Rutuja Nandre¹, Hyerang Eom¹

¹Department of BioMedical Bigdata (BK21) and Research Institute of Life Sciences, Gyeongsang National University, Jinju 52828, Republic of Korea.

Journal of microbiology (Seoul, Korea)

|July 2, 2025

概括

研究人员开发了一种基因编辑热友菌,Paecilomyces variotii,用于增强蛋白质生产. 他们成功地使用核糖蛋白 (RNP) 介导的基因编辑来创建具有改善异质蛋白表达的菌株,为工业应用铺平了道路.

科学领域:

生物技术是生物技术.
分子生物学分子生物学
菌类学菌类学是指菌类学.

背景情况:

热性真菌为工业蛋白质生产提供了优势.
帕西洛米切斯 (Paecilomyces variotii) 是一种热性真菌,具有作为蛋白质生产宿主的潜力.
开发高效的基因编辑工具对于优化真菌宿主至关重要.

研究的目的:

在Paecilomyces variotii中建立基因编辑系统,用于异构蛋白质的生产.
为了安全处理和环境封闭,创建一个缺乏状菌株.
为了评估不同的促进剂强度和信号序列对蛋白质表达的影响.

主要方法:

隔离和表征一个热性Paecilomyces variotii菌株 (MR1).
紫外线突变产生一种缺乏菌的菌株 (UM7).
Cas9-gRNA核糖核蛋白 (RNP) 介导的基因编辑与同质导向修复 (HDR) 使用pyrG作为选择标记.
原生体与含有各种促进子序列和eGFP的RNP和HR捐赠者DNA共同转化.
序列分析以确认成功的HDR和推广器效率的评估.

主要成果:

成功生成了P. variotii. 的一个缺乏状菌株 (UM7) 的菌株.
在P. variotii中展示了RNP介导的基因编辑和HDR,产生了16个具有整合促进体-eGFP结构的pyrG破坏突变.
确定PgpdA是细胞内GFP生产最强的促进剂.
观察信号对细胞外GFP表达产生负面影响.
增强GFP表达在细胞和培养中,在一个新编辑的菌株中缺乏信号序列.

结论:

这项研究报告了第一个成功的RNP介导基因编辑在Paecilomyces variotii.
开发的基因编辑系统可以优化P. variotii作为异质蛋白质生产的主体.
消除信号显著改善了蛋白质表达和分泌,突出了其在优化真菌蛋白质生产平台中的作用.

关键词:

这就是HDR HDR.这种植物是Paecilomyces variotii.在 RNP RNP 中.基因编辑基因编辑不同类型的蛋白质.

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