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Updated: Jan 18, 2026

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顺序的因子传递使得在临床级的iPS细胞中实现通用基因编辑的高效工作流

Thomas Berger¹, Elitsa Borisova¹, Anna Gamerschlag¹

¹Catalent Duesseldorf GmbH, Berghausener Straße 98, 40764, Langenfeld, Germany.

Scientific reports

|September 12, 2025

概括

我们优化了CRISPR基因编辑用于人类诱导多能干细胞 (iPSCs),为细胞治疗应用实现了超过30%的敲门效率. 这种无病毒的方法提高了治疗用途的安全性和免疫兼容性.

科学领域:

干细胞生物学干细胞生物学
基因编辑技术的技术
细胞疗法细胞疗法

背景情况:

人类诱导的多能干细胞 (iPSCs) 对细胞治疗有很大的前景.
克里斯普尔技术可以在iPSC中进行基因修改,以增强治疗功能.
目前对iPSCs的基因操纵,特别是敲进,是低效的,可能与良好制造实践 (GMP) 相冲突.

研究的目的:

优化基于CRISPR的基因编辑程序,以在iPSC中有效地产生内置基因.
开发一种符合GMP的,无病毒的方法来对iPSC进行基因操纵.
创建和验证具有增强免疫兼容性和安全功能的iPSC线.

主要方法:

基于Cas9和Cas12a的核糖蛋白 (RNP) 复合物的系统优化,用于基因编辑.
连续交付RNP和捐赠体等离子体,以免病毒敲进.
创建和表征同卵性iPSC线条与HLA类I耗尽和可诱导的caspase-9自杀基因.

主要成果:

使用优化方法实现了全长转基因的超过30%的连续效率.
成功生成了缺乏HLA类I和表达安全开关的同卵性iPSC线.
在GMP iPSC系列中验证了基因组完整性,差异化能力和安全开关功能.

结论:

改进的基因编辑方法为iPSC修改提供了灵活和高效的平台.
这种无病毒的方法克服了与当前基因编辑技术相关的GMP限制.
开发的iPSC系列适用于基础研究,并有可能用于细胞治疗应用.

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