Hedgehog (Hh) Reporter Activity Assay

Abstract

This protocol is for testing responses of a candidate cell line/cell lines to Hh ligands or Hh pathway agonists stimulation. This protocol can also be adapted to screen small molecule libraries or biologics that contain activities to either increase or decrease Hh pathway responses. Canonical Hh signaling activity transcriptionally induces Hh target genes that contain consensus Gli binding element. Hh-responsive cells transiently or stably expressing luciferase protein under the regulation of the Gli promoter element can be used to report stimulus-dependent Hh-pathway activity., 该方案用于测试候选细胞系/细胞系对Hh配体或Hh通路激动剂刺激的反应。 该方案还可以适于筛选含有增加或降低Hh通路反应的活性的小分子文库或生物制剂。 规范Hh信号活动转录诱导含有共有Gli结合元件的Hh靶基因。 在Gli启动子元件的调节下瞬时或稳定表达荧光素酶蛋白的Hh反应性细胞可用于报告刺激依赖性Hh-途径活性。