Visual barcodes for clonal-multiplexing of live microscopy-based assays

• DOI: 10.1038/s41467-022-30008-0
• Publication Date: 2022-05-18T10:02:54Z
• Authors: Tom Kaufman; Erez Nitzan; Nir Firestein; Miriam Bracha Gin...; Seshu Iyengar; Nish Patel; Rotem Ben-Hamo; Ziv Porat; Jaryd Hunter; Andreas Hilfinger; Varda Rotter; Ran Kafri; Ravid Straussman
• Keywords: Microscopy; 0303 health sciences; 03 medical and health sciences; Science; Q; DNA Barcoding, Taxonomic; DNA; Article; Clone Cells

Abstract While multiplexing samples using DNA barcoding revolutionized the pace of biomedical discovery, live imaging-based applications has been limited by number fluorescent proteins that can be deconvoluted common microscopy equipment. To address this limitation, we develop visual barcodes discriminate clonal identity single cells different are targeted to specific subcellular locations. We demonstrate deconvolution these is highly accurate and robust many cellular perturbations. then use generate ‘Signalome’ cell-lines mixing 12 clones reporters into a population, allowing simultaneous monitoring activity in branches signaling, at resolution, over time. Using identify two distinct clusters signaling pathways balance growth proliferation, emphasizing importance homeostasis as central organizing principle cancer signaling. The ability multiplex imaging applications, both vitro vivo may allow better high-content characterization complex biological systems.