Novartis Institutes for BioMedical Research, Cambridge, MA, USA.
Novartis Institutes for BioMedical Research, Basel, Switzerland.
Sci Rep. 2020 Feb 7;10(1):2097. doi: 10.1038/s41598-020-58861-3.
Gene and compound functions are often interrogated by perturbation. However, we have limited methods to capture associated phenotypes in an unbiased and holistic manner. Here, we describe Fluopack screening as a novel platform enabling the profiling of subcellular phenotypes associated with perturbation. Our approach leverages imaging of a panel of fluorescent chemical probes to survey cellular processes in an unbiased and high throughput fashion. Segmentation-free, whole image analysis applied to Fluopack images identifies probes revealing distinct phenotypes upon perturbation, thereby informing on the function and mechanism of action of perturbagens. This chemical biology approach allows to interrogate phenotypes that tend to be overlooked by other methods, such as lipid trafficking and ion concentration inside the cell. Fluopack screening is a powerful approach to study orphan protein function, as exemplified by the characterization of TMEM41B as novel regulator of lipid mobilization.
基因和化合物的功能通常通过干扰来进行研究。然而，我们只有有限的方法能够以无偏倚和全面的方式获取相关表型。在这里，我们描述了 Fluopack 筛选，这是一种新的平台，可以对与干扰相关的亚细胞表型进行分析。我们的方法利用一组荧光化学探针的成像来以无偏倚和高通量的方式研究细胞过程。应用于 Fluopack 图像的无分割、全图像分析可以识别出在受到干扰时显示出不同表型的探针，从而提供关于干扰剂的功能和作用机制的信息。这种化学生物学方法可以研究其他方法容易忽略的表型，例如细胞内的脂质转运和离子浓度。Fluopack 筛选是研究孤儿蛋白功能的一种强大方法，例如将 TMEM41B 鉴定为新型脂质动员调节剂。
