Luciferase assay system to monitor fibroblast growth factor signal disruption in human iPSCs

STAR Protoc. 2022 Jun 4;3(2):101439. doi: 10.1016/j.xpro.2022.101439. eCollection 2022 Jun 17.

Abstract

We describe a protocol for a live-cell luciferase assay system for continuously monitoring fibroblast growth factor (FGF) signal disruption in human-induced pluripotent stem cells (iPSCs). Signal disrupting effects of chemicals are used as an indicator to evaluate toxicity. The assay is reliably predictive of the effects of limb malformation chemicals (AUC = 0.93). The current approach is limited to FGF signal disruption, and combinations with other types of signaling will be required to detect the effects of different toxicants. For complete details on the use and execution of this protocol, please refer to Kanno et al. (2022a).

Keywords: Biotechnology and bioengineering; CRISPR; Cell Biology; Cell-based Assays; Molecular Biology; Stem Cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fibroblast Growth Factors / metabolism
  • Humans
  • Induced Pluripotent Stem Cells* / metabolism
  • Luciferases / genetics
  • Signal Transduction

Substances

  • Fibroblast Growth Factors
  • Luciferases