Shin Young-Hee, Park Seung Bum
Department of Chemistry, Seoul National University, Seoul, South Korea.
Department of Biophysics and Chemical Biology, Seoul National University, Seoul, South Korea.
Methods Mol Biol. 2018;1787:223-234. doi: 10.1007/978-1-4939-7847-2_17.
Phenotypic screening in live cells has emerged as a promising strategy for drug discovery in pharmaceutical communities. For relevant phenotype-based screening setups, it is critical to develop adequate reporters in order to selectively visualize subcellular compartments or phenotypic changes that represent disease-related characteristics during compound screening. In this chapter, we introduce two phenotype-based high-content/high-throughput assays using fluorescent bioprobes that have been designed and refined to selectively stain cellular lipid droplets (LDs) and to show cellular glucose uptake. In conjunction with target identification process for the hit compounds from phenotypic screening, these fluorescent chemical probe-based screening techniques are expected to drive a great advancement for the discovery of novel first-in-class therapeutics.
活细胞中的表型筛选已成为制药界药物发现的一种有前景的策略。对于相关的基于表型的筛选设置，开发合适的报告分子至关重要，以便在化合物筛选过程中选择性地可视化代表疾病相关特征的亚细胞区室或表型变化。在本章中，我们介绍了两种基于表型的高内涵/高通量分析方法，这些方法使用了经过设计和优化的荧光生物探针，以选择性地染色细胞脂滴（LDs）并显示细胞对葡萄糖的摄取。结合从表型筛选中获得的命中化合物的靶点识别过程，这些基于荧光化学探针的筛选技术有望推动新型一流疗法的发现取得巨大进展。
