可通过CRISPR或shRNA生成内源性逆转录病毒样颗粒缺乏的CHO细胞,并根据逆转录病毒包膜蛋白的表达进行丰富
Matthew Stuible1, Sergio P Alpuche-Lazcano1,2, Christian Gervais1
1National Research Council Canada, Human Health Therapeutics Research Centre, Montreal, Quebec, Canada.
Biotechnology and bioengineering
|August 22, 2025
概括
克里斯普尔和shRNA基因编辑有效地降低了中国仓鼠卵巢 (CHO) 细胞的逆转录病毒样粒子 (RVLP) 生产. 这提高了生物制造的安全性,通过减轻与RVLP污染相关的风险.
科学领域:
- 生物技术
- 分子生物学
- 细胞生物学
背景情况:
- 来自中国仓鼠卵巢 (CHO) 细胞内源序列的复原病毒样颗粒 (RVLP) 在生物制造中构成安全风险.
- 尽管缺乏传染性,但RVLP与哺乳动物白血病病毒相似,需要采取缓解策略.
研究的目的:
- 开发和验证用于破坏CHO细胞中的RVLP生产的基因工程策略.
- 建立用于生物制造应用的RVLP缺乏的CHO细胞克隆的方法.
主要方法:
- 使用CRISPR和shRNA基因编辑以准RVLP核酸序列.
- 在CHO细胞表面检测RVLP包膜 (Env) 特定抗体.
- 根据减少RVLP蛋白表达选择RVLP缺乏克隆.
主要成果:
- 在工程化CHO细胞中,CRISPR和shRNA有效降低了RVLP的产生.
- 基于抗体的查有助于有效选择缺乏RVLP的克隆.
- 修改后的CHO细胞维持或增强了复合病毒样粒子 (VLP) 的产生.
结论:
- 基因工程提供了一种可行的策略,以减轻CHO细胞衍生生物制造中RVLP相关的安全风险.
- 这种方法对于与传统病毒清除方法不相容的复杂生物药物特别有益.
- 缺乏RVLP的CHO细胞为生产生物治疗药物提供了更安全的平台.
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