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Updated: Jun 5, 2025

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Published on: June 16, 2017

开发一种RecT辅助的内源CRISPR/SzCas9系统,用于精确编辑Streptococcus zooepidemicus的基因组

Xuejing Zhang¹, Feiyu Wang¹, Xueping Guo²

¹State Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, People's Republic of China.

International journal of biological macromolecules

|December 14, 2024

概括

研究人员开发了一种新的CRISPR基因编辑系统,用于Streptococcus zooepidemicus,改善了氨酸的产生. 该系统可实现精确的基因改造,从而提高产量,并为工业应用提供更安全的产品.

科学领域:

微生物学微生物学
生物技术是生物技术.
基因工程是一种基因工程.

背景情况:

Streptococcus zooepidemicus 是工业化氨酸 (HA) 生产的关键.
有限的遗传工具阻碍了用于HA制造的S. zooepidemicus菌株改进.

研究的目的:

开发一种高效,无标记的基因编辑系统,用于工业S. zooepidemicus菌株.
为了改造S. zooepidemicus以提高氨酸的产量和提高安全性,

主要方法:

为S. zooepidemicus开发了一种RECT辅助的内源CRISPR/SzCas9系统.
进行了无标记基因删除 (hylb),基因替代 (VHb) 和停止编码子插入 (sagA).
使用代编辑来实现所需的遗传修改.

主要成果:

成功设计了一种具有降低毒性的非溶血性S. zooepidemicus菌株 (SD-3).
在 Hyaluronic 酸产量上实现了 39.7% 的增长,达到 10.8 g/L.
生产的高分子量 hialuronic 酸 (高达 2.8 x 10^6 Da) 具有统一的特性.

结论:

新的CRISPR/SzCas9系统为S. zooepidemicus提供了高效的基因操纵.
改造后的SD-3菌株显示出工业化氨酸生产的巨大潜力,提高了产量和安全性.

关键词:

这就是CRIPSR/Cas9的原因.基因组编辑基因组编辑高分子量氨酸是什么?代谢工程是代谢工程.Streptococcus zooepidemicus 这种细菌是动物性流行病.

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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

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

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