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Updated: May 29, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

在Ganoderma lucidum的二核菌株中同时对两核进行基因编辑,使用Cas9-gRNA核糖蛋白

Yeon-Jae Choi¹, Hyerang Eom¹, Rutuja Nandre¹

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

Journal of microbiology (Seoul, Korea)

|February 3, 2025

概括

现在可以使用Cas9-gRNA核糖蛋白复合体 (RNP) 在核中进行基因编辑. 这种方法成功地修改了Ganoderma lucidum的两个核中的pyrG基因,克服了基因修改方面的挑战.

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
基因工程是一种基因工程.

背景情况:

多核真菌细胞对遗传修饰提出了挑战.
针对共享细胞质内的多个核中的特定基因是复杂的.

研究的目的:

为了证明二核生物真菌的两个核中的成功基因编辑.
为了利用Cas9-gRNA核糖核蛋白复合体 (RNP) 在Ganoderma lucidum中进行基因修饰.

主要方法:

引入针对pyrG基因的Cas9-gRNA RNP,进入二核生物的G. lucidum原生体.
对抗5-酸 (5-FOA) 的变压剂的选择.
使用交配类型标记物和pyrG基因中断的序列分析确认了二核细胞菌株.

主要成果:

隔离了31个耐药菌株,其中26个被确认为二核菌株.
在两个核中 (pyrG-/pyrG-) 或在一个核中 (pyrG+/pyrG-或pyrG-/pyrG+) 发现具有pyrG破坏的二.
在6个月内通过非同类末端连接修复证明稳定的核基因编辑.

结论:

Cas9-gRNA RNP 能够在一个共同的细胞质中跨越多个核进行高效的基因编辑.
这一突破促进了诸如Ganoderma lucidum之类的多核真菌的遗传修饰.
该方法显示了在真菌研究和应用中稳定基因操纵的潜力.

关键词:

卡尔基奥斯基的卡尔基奥斯基是什么意思基因编辑基因编辑异体菌症的异体菌症是什么是一种真菌.

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