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结合单细胞CRISPR查和表观基因分析揭示了因果基因调控网络
Adam J Rubin1, Kevin R Parker2, Ansuman T Satpathy3
1Program in Epithelial Biology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Cell
|December 25, 2018
概括
Perturb-ATAC是一种新的单细胞方法,通过将CRISPR基因编辑与染色体可访问性分析相结合来绘制基因调节图. 这种方法揭示了转录因子如何控制人体细胞的身份和发育.
科学领域:
- 基因组学
- 分子生物学
- 细胞生物学
背景情况:
- 了解基因调节网络对于解读细胞发育和疾病至关重要.
- 目前的方法往往不足以剖析单个细胞内的复杂相互作用.
研究的目的:
- 介绍Perturb-ATAC,一种用于基因调控网络的新型单细胞技术.
- 研究转录因子,染色体修饰剂和非编码RNA在细胞过程中的作用.
主要方法:
- Perturb-ATAC结合了多重的CRISPR干扰/敲击与单细胞ATAC-seq.
- 同时检测导向RNA和开放的染色体位点可以进行基因型-表型映射.
- 应用于约4,300个单细胞,分析了63个以上的基因型-表型关系.
主要成果:
- 在人类B淋巴细胞中,确定了染色质可访问性,TF占用率和核细胞定位的调节剂.
- 发现了管理B细胞状态,变异和疾病相关元素的TF层次结构.
- 在表皮细胞中,揭示了测定质细胞命运的序列 cis 元素模块.
- 发现基于基因组共同定位的TF表观关系和协同作用.
结论:
- Perturb-ATAC是解剖基因调节网络的强大策略.
- 这种方法为细胞决策过程提供了前所未有的洞察力.
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