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人类基因组重写用于现成的干细胞揭示了"表观遗传鬼魂"
Serena F Generoso1, Sarah Levovitz1, Susanna Jaramillo1
1Department of Biomedical Engineering, NYU Tandon School of Engineering, Brooklyn, NY, 11201 USA.
bioRxiv : the preprint server for biology
|September 26, 2025
概括
我们设计了一个新的基因组编写平台REWRITE,用于修改干细胞中的人类白细胞抗原 (HLA) 基因. 这一突破可以通过克服免疫排斥来开发通用,现成的细胞疗法.
科学领域:
- 基因组学和基因编辑
- 免疫学和移植
- 干细胞生物学 干细胞生物学
背景情况:
- 人类白细胞抗原 (HLA) 基因多样性是创建通用兼容细胞疗法的重要障碍.
- 现有的基因组编辑技术在高效修改复杂基因工程所需的大型DNA段方面存在局限性.
研究的目的:
- 开发一种用于人类多能干细胞 (hPSCs) 大规模精确基因组编辑的新平台.
- 用修改的HLA位点来设计hPSC,以克服对全原细胞疗法的免疫排斥.
- 研究与大规模遗传修饰相关的表观遗传现象.
主要方法:
- 开发REWRITE,一个模块化,减少痕的平台,用于对大型合成结构 (>100 kb) 的代基因组编写.
- 应用REWRITE来删除内源HLA位点的大量部分,并在hPSC中安装合成HLA单元型和抗原处理基因.
- 对表观遗传状态的分析,包括对"表观遗传鬼魂"的识别,以及对差异化细胞系中的基因表达和免疫兼容性的评估.
主要成果:
- 成功删除了HLA位点的105-209kb,并集成了高达100kb的合成结构.
- 发现了一种持久的"表观遗传幽灵"现象,在遗传删除后,活跃状态会持续存在,这由本地DNA元素来解决.
- 工程细胞显示恢复诱导表达,抵抗自然杀手 (NK) 细胞中介杀伤,并建立了T细胞耐受性,这对于现成疗法至关重要.
结论:
- 该REWRITE平台能够在hPSC中实现高效的大规模基因组工程,克服HLA多态性挑战.
- 这些发现揭示了对表观遗传记忆及其在基因组编辑过程中的分辨率的新见解.
- 这项技术有助于创建"现成"的全源细胞疗法,并为更广泛的基因组编程应用提供了潜力.
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