FRET and BRET-based biosensors in live cell compound screens

Katie Herbst Robinson, Jessica R. Yang, Jin Zhang

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Live cell compound screening with genetically encoded fluorescence or bioluminescence-based biosensors offers a potentially powerful approach to identify novel regulators of a signaling event of interest. In particular, compound screening in living cells has the added benefit that the entire signaling network remains intact, and thus the screen is not just against a single molecule of interest but against any molecule within the signaling network that may modulate the distinct signaling event reported by the biosensor in use. Furthermore, only molecules that are cell permeable or act at cell surface receptors will be identified as "hits," thus reducing further optimization of the compound in terms of cell penetration. Here we discuss a detailed protocol for using genetically encoded biosensors in living cells in a 96-well format for the execution of high throughput compound screens and the identification of small molecules which modulate a signaling event of interest.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages217-225
Number of pages9
Volume1071
ISBN (Print)9781627036214
DOIs
StatePublished - 2014

Publication series

NameMethods in Molecular Biology
Volume1071
ISSN (Print)10643745

Keywords

  • Bioluminescence
  • Cell signaling
  • Fluorescence resonance energy transfer
  • Genetically encoded biosensor
  • Live cell compound screening

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Robinson, K. H., Yang, J. R., & Zhang, J. (2014). FRET and BRET-based biosensors in live cell compound screens. In Methods in Molecular Biology (Vol. 1071, pp. 217-225). (Methods in Molecular Biology; Vol. 1071). Humana Press Inc.. https://doi.org/10.1007/978-1-62703-622-1_17