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人体细胞中的光学聚合屏幕
David Feldman1, Avtar Singh2, Jonathan L Schmid-Burgk2
1Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Department of Physics, MIT, Cambridge, MA 02142, USA.
Cell
|October 19, 2019
概括
这项研究引入了哺乳动物细胞的聚合光学遗传查方法,使复杂细胞过程的基于图像的表型化成为可能. 这种新方法成功地确定了参与核因子-卡帕B (NF-κB) 信号动态的基因.
科学领域:
- 细胞生物学
- 遗传学
- 分子生物学
背景情况:
- 基因选对于识别控制细胞表型的基因至关重要.
- 聚合基因屏增强了可扩展性,但与动态细胞行为的详细成像不相容.
- 现有的方法缺乏高吞吐量,基于图像的聚合遗传库选能力.
研究的目的:
- 为哺乳动物细胞开发一个聚合光学基因查方法,该方法将基于图像的表型与聚合图书馆解复合结合起来.
- 应用这种方法来识别参与核因子-kappa B (NF-κB) 信号传递的基因.
- 使用活细胞成像研究NF-κB信号的时间调节.
主要方法:
- 开发了一种使用针对性 in situ 测序进行解复的聚合基因查方法.
- 通过对RelA (p65) 核转位进行成像,对数百万个细胞中的952个基因进行了选.
- 使用活细胞成像来捕捉p65核保留的动态变化.
主要成果:
- 在三个细胞系的单一时间点屏幕中确定了NF-κB途径的15个已知组成部分.
- 通过活细胞成像来调节p65核保留的时间.
- 证明了使用聚合图书馆选空间和时间定义的表型的能力.
结论:
- 开发的聚合光学基因查方法使得哺乳动物细胞聚合图书馆的基于图像的高通量表型化成为可能.
- 这种方法有效地识别信号通路的组件并剖析它们的时间动态.
- 这种方法为研究复杂的细胞表型提供了前所未有的规模和分辨率的强大工具.
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