Fluorescent genetic barcoding in mammalian cells for enhanced multiplexing capabilities in flow cytometry

摘要: The discovery of the green fluorescent protein from Aequorea victoria has revolutionized field cell and molecular biology. Since its a growing panel proteins, fluorophores fluorescent-coupled staining methodologies, have expanded analytical capabilities flow cytometry. Here, we exploit power genetic engineering to barcode individual cells with genes encoding proteins. For engineering, utilize retroviral technology, which allows for expression ectopic information in stable manner mammalian cells. We genetically barcoded both adherent nonadherent different Multiplexing was increased by combining number distinct fluorescence intensity each channel. Moreover, proven be at least 6-month period, is critical applications such as biological screens. shown applicability multiplexing cell-based assays that rely themselves on barcoding, or classical protocols. Fluorescent barcoding gives an inherited characteristic distinguishes it counterpart. Once lines are developed, no further manipulation required, decreasing time, nonspecific background associated protocols, cost. increasing discovered and/or engineered proteins unique absorbance/emission spectra, combined detection devices lasers, increases versatility, making powerful tool cytometry-based analysis.