Multiplexable fluorescence lifetime imaging (FLIM) probes for Abl and Src-family kinases. Issue 87 (9th October 2020) - Jena, Sampreeti
Abstract : Here we describe development of cell-penetrating peptide substrates for fluorescence lifetime imaging (FLIM) of Abl and Src-family kinase activities, applied to mapping single cell and subcellular dynamics of multiple kinases in live cells. Abstract : Many commonly employed strategies to map kinase activities in live cells require expression of genetically encoded proteins ( e.g. FRET sensors). In this work, we describe the development and preliminary application of a set of cell-penetrating, fluorophore labelled peptide substrates for fluorescence lifetime imaging (FLIM) of Abl and Src-family kinase activities. These probes do not rely on FRET pairs or genetically-encoded protein expression. We further demonstrate probe multiplexing and pixel-by-pixel quantification to estimate the relative proportion of modified probe, suggesting that this strategy will be useful for detailed mapping of single cell and subcellular dynamics of multiple kinases concurrently in live cells.