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通过Tn5转换和在组织中确定性DNA条形编码进行空间解析的表观基因组测序
Negin Farzad1,2, Archibald Enninful1,2, Shuozhen Bao1,2
1Department of Biomedical Engineering, Yale University, New Haven, CT, USA.
Nature protocols
|June 28, 2024
概括
本研究介绍了两个空间表观基因组分析方法,即空间CUT&Tag和空间ATAC测序,用于详细的组织分析. 这些技术可以在原生组织环境中对表观遗传景观进行公正的,高分辨率的绘制.
科学领域:
- 表观遗传学 在表观遗传学中,表观遗传学是指表观遗传学.
- 基因组学就是基因组学.
- 分子生物学分子生物学
背景情况:
- 空间表观遗传映射对于理解局部组织环境中的基因调节至关重要.
- 现有的方法可能需要对特定标记物的预先了解,从而限制了公正的探索.
- 需要高分辨率的空间分析来捕捉取决于组织背景的细胞功能.
研究的目的:
- 为两个新的空间表观基因组分析方法提供一个全面的协议:空间-CUT&Tag和空间-ATAC-测序.
- 为了实现全基因组,无偏见的表观遗传特征在一个像素一点的水平,单基底分辨率.
- 为各种物种和组织促进这些技术的适应.
主要方法:
- 开发用于基因组位置识别的组织内Tn5转移.
- 集成微流体确定性条形码用于空间地址编码.
- 建立了用于样本准备,图书馆构建,测序和数据分析的协议.
主要成果:
- 空间CUT&Tag的可行性已被证明,用于组织蛋白修饰概况.
- 展示了用于染色体可访问性映射的空间ATAC测序.
- 实现了10μm像素大小分辨率和单基分辨率用于表观遗传景观.
- 为数据解释提供定制分析管道.
结论:
- 空间表观遗传学分析方法,如空间CUT&Tag和空间ATAC测序,为研究组织特异性表观遗传学调节提供了强大的工具.
- 这些方法可以适应不同物种,不需要先前的标记知识.
- 该协议是高效的,可以在2-3天内完成四个样本,并且可以通过GitHub访问.
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