OAZ1 / CASP8AP2 双淘汰赛通过代谢重编程和抗亡作用增强了HEK293细胞中重组蛋白质的产生
Junhe Zhang1,2,3, Liao Zhang1,2, Lu Hou1,2
1Institutes of Health Central Plains, Xinxiang Key Laboratory for Tumor Drug Screening and Targeted Therapy, Henan Medical University, Xinxiang 453003, China.
Acta biochimica et biophysica Sinica
|January 28, 2026
概括
我们通过敲除OAZ1和CASP8AP2基因,增强了HEK293细胞中的重组蛋白质生产,减少了细胞亡,并优化了新陈代谢以获得更高的产量.
科学领域:
- 生物技术是生物技术.
- 细胞生物学 细胞生物学
- 分子生物学分子生物学
背景情况:
- 人类胚胎 (HEK) 293细胞由于它们的翻译后修饰能力,对重组蛋白质生产至关重要.
- 大规模的HEK293细胞培养面临由代谢压力和亡造成的限制,减少蛋白质产量.
研究的目的:
- 通过调节代谢和亡途径,增强HEK293细胞中的重组蛋白表达.
- 为了创建一个双击HEK293细胞系,针对OAZ1和CASP8AP2基因.
主要方法:
- 利用CRISPR/Cas9技术,淘汰了ornithine脱碳酶抗酶1 (OAZ1) 和酶8相关蛋白2 (CASP8AP2) 基因.
- 通过单细胞克隆和遗传验证构建并验证了一个OAZ1/CASP8AP2双击HEK293细胞系.
- 进行了代谢分析和评估了重组蛋白质生产 (SEAP, VN).
主要成果:
- 成功生成了一个OAZ1/CASP8AP2双淘汰的HEK293细胞系,确认了基因淘汰.
- 与野生类型相比,在双击的细胞中观察到显著降低了亡率.
- 实现了SEAP的2.1倍增长和越南生产的2.9倍增长.
- 确定了G1/G0细胞循环停止和增加了特定代谢物消耗/生产率.
结论:
- 同时抑制亡和代谢优化有效地增强了HEK293细胞中的重组蛋白质生产.
- OAZ1/CASP8AP2双淘汰策略为改善HEK293基于细胞系统中的蛋白质表达提供了一种新的方法.
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