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单细胞和早期胚胎中的多能性因子分析
Sarah J Hainer1, Ana Bošković2, Kurtis N McCannell3
1Department of Molecular, Cell, and Cancer Biology, University of Massachusetts Medical School, Worcester, MA 01605, USA; Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260, USA.
Cell
|April 9, 2019
概括
我们开发了超低输入的CUT&RUN (uliCUT&RUN) 来映射单细胞中的转录因子 (TF) 结合部位. 这种方法揭示了TF结合的动态和依赖性,对于理解细胞在发育和疾病中的命运决定至关重要.
科学领域:
- 发育生物学
- 基因组学
- 分子生物学
背景情况:
- 细胞命运决定依赖于在小细胞群中起作用的转录因子 (TF).
- 在单细胞水平上在体内识别TF结合点是当前技术的挑战.
研究的目的:
- 调整CUT&RUN试验以分析低细胞数,包括单细胞的转录因子.
- 研究罕见细胞群中的TF结合位点占用率和依赖性.
主要方法:
- 通过核酶 (CUT&RUN) 试验对超低输入 (uliCUT&RUN) 的目标和释放进行裂变的调整.
- 应用uliCUT&RUN在单细胞和个体植入前胚胎中的转录因子.
主要成果:
- 单细胞实验表明大多数细胞中只占据了TF结合部位的一小部分.
- 多能性TF NANOG的染色体结合高度依赖于单个细胞囊中的SWI/ SNF染色体重塑复合物.
结论:
- 超低输入的CUT&RUN (uliCUT&RUN) 能够在罕见细胞群中实现全基因组TF结合分析.
- 这种技术对于研究发育和疾病中的TF功能,特别是在胚胎环境中是有价值的.
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