Skip to Main content Skip to Navigation
Journal articles

Automated screening of AURKA activity based on a genetically encoded FRET biosensor using fluorescence lifetime imaging microscopy

Abstract : Fluorescence Lifetime Imaging Microscopy (FLIM) is a robust tool to measure Förster Resonance Energy Transfer (FRET) between two fluorescent proteins, mainly when using genetically-encoded FRET biosensors. It is then possible to monitor biological processes such as kinase activity with a good spatiotemporal resolution and accuracy. Therefore, it is of interest to improve this methodology for future high content screening purposes. We here implement a time-gated FLIM microscope that can image and quantify fluorescence lifetime with a higher speed than conventional techniques such as Time-Correlated Single Photon Counting (TCSPC). We then improve our system to perform automatic screen analysis in a 96-well plate format. Moreover, we use a FRET biosensor of AURKA activity, a mitotic kinase involved in several epithelial cancers. Our results show that our system is suitable to measure FRET within our biosensor paving the way to the screening of novel compounds, potentially allowing to find new inhibitors of AURKA activity.
Document type :
Journal articles
Complete list of metadatas

Cited literature [1 references]  Display  Hide  Download

https://hal-univ-rennes1.archives-ouvertes.fr/hal-02500623
Contributor : Xavier Chard-Hutchinson <>
Submitted on : Tuesday, March 17, 2020 - 4:47:58 PM
Last modification on : Thursday, March 19, 2020 - 9:09:11 AM
Long-term archiving on: : Thursday, June 18, 2020 - 3:06:17 PM

File

Sizaire et al-2020-Automated s...
Files produced by the author(s)

Identifiers

Citation

Florian Sizaire, Gilles Le Marchand, Jacques Pécréaux, Otmane Bouchareb, Marc Tramier. Automated screening of AURKA activity based on a genetically encoded FRET biosensor using fluorescence lifetime imaging microscopy. Methods and Applications in Fluorescence, IOP Science, 2020, 8 (2), pp.024006. ⟨10.1088/2050-6120/ab73f5⟩. ⟨hal-02500623⟩

Share

Metrics

Record views

271

Files downloads

50