相关实验视频
Updated: Feb 10, 2026

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Genome Editing in Mammalian Cell Lines using CRISPR-Cas
Published on: April 11, 2019
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使用Cas-CLOVER系统评估CHO细胞中的基因编辑
Tiffany McLamarrah1, Efecan Aral1, Michael Hoffman1
1Mammalian Platform, Global CMC Development, Framingham, Massachusetts, USA.
Biotechnology progress
|February 8, 2026
概括
使用Cas-CLOVER的基因编辑精确地准了中国仓鼠卵巢 (CHO) 细胞中的谷氨酸合成酶 (GS) 位点. 这导致单克隆抗体产生显著增强,显示标位增加了14.5倍.
科学领域:
- 生物技术是生物技术.
- 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:
- 中国仓鼠卵巢 (CHO) 主体对于生物制药生产至关重要.
- 基因编辑技术为宿主细胞系发展提供了先进的方法.
- 提高CHO电池的稳定性和生产率对于高效的制造至关重要.
研究的目的:
- 在CHO细胞中使用Cas-CLOVER可编程核酶进行精确的基因编辑.
- 为了生成和验证Glutamine合成酶淘汰 (GSKO) CHO宿主细胞系.
- 评估GS基因淘汰对重组蛋白质生产的影响.
主要方法:
- 使用Cas-CLOVER可编程核酶来准CHO-K1细胞中的谷氨酸合成酶 (GS) 位点.
- 从30个独特的细胞系产生了100多个潜在的GS淘汰 (GSKO) 克隆.
- 用三种正交法方法验证GSKO克隆,以确认基因淘汰.
主要成果:
- 在生成的GSKO克隆中,在98个克隆中确认了GS基因的完全淘汰.
- 使用精选的GSKO克隆进行单克隆抗体生产,显示出提高了生产率.
- 与野生类型的父母宿主相比,观察到标位的显著增加,高达14.5倍.
结论:
- Cas-CLOVER介导的基因编辑是开发改进的CHO宿主细胞系的有效工具.
- 在GSKO CHO细胞系中,重组蛋白质生产的生产率显著提高.
- 这种基因编辑方法在优化生物制药制造工艺方面具有巨大的潜力.
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