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新生转录组的动力学和空间基因组学 by Intron seqFISH
Sheel Shah1, Yodai Takei2, Wen Zhou2
1Division of Biology and Biological Engineering, Caltech, Pasadena, CA 91125, USA; UCLA-Caltech Medical Scientist Training Program, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
Cell
|June 12, 2018
概括
通过现场光杂交 (seqFISH) 进行内部测序,可在单细胞中可视化新生的RNA转录. 这种方法揭示了小鼠细胞中的动态核组织和异步基因表达振荡.
科学领域:
- 分子生物学
- 基因组学
- 细胞生物学
背景情况:
- 在单个细胞中可视化转录组和核组织仍然是一个重大挑战.
- 现有的方法很难在现场捕捉活跃的转录点的动态.
研究的目的:
- 开发一种新的多重化方法,在现场对新生转录组进行单分子可视化.
- 研究单个细胞内的活跃转录位点的空间组织和动态.
- 使用新生转录组概况识别细胞类型和状态.
主要方法:
- 通过现场光混合 (intron seqFISH) 开发了内测序,这是一个多重化单分子的现场方法.
- 在新生转录活性位点启用了10421个基因的成像.
- 结合mRNA和lncRNAseqFISH和免疫光学进行综合分析.
主要成果:
- 在小鼠胚胎干细胞和纤维细胞中成功识别了不同的细胞类型和状态.
- 证明新生RNA合成部位通常位于具有高细胞间变异性的染色体区域表面.
- 在两种细胞类型中发现全球新生转录的异步振荡.
结论:
- Intron seqFISH提供了前所未有的单细胞新生转录组的空间基因组学.
- 揭示了核组织的基本原理和以前模糊的快速转录动态.
- 为剖析细胞异质性和基因调控提供了强大的工具.
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