Gene Transfer in J-774 cells with lipid-based reagents and use of GFP as a marker in flow cytometry measurements
DNA transfection is a widely used tool for the study of gene expression and mechanisms of protein function. Macrophagic cells are refractory to efficient transfection, owing most probably to their specific physiology. An efficient gene transfer technique may nevertheless be prerequisite for the stud...
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[Yogyakarta] : Universitas Gadjah Mada
2001
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| Summary: | DNA transfection is a widely used tool for the study of gene expression and mechanisms of protein function. Macrophagic cells are refractory to efficient transfection, owing most probably to their specific physiology. An efficient gene transfer technique may nevertheless be prerequisite for the study of their macrophage functions. We describe in this paper comparative studies of transfection using commercially available reagents. We used J-774 cell line as it already exhibits caracteristics of macrophage without a prior differentiation process. Not all reagents can be used to successfully transfect J-774 cells. With the most efficient reagents, up to 14% of cells were transfected, using GFP as reporter. Due to the autofluorescence of GFP, transfection efficiencies were directly and easily measured by flow cytometry
Keywords: flow cytometry - gene transfer - GFP - J-774 - transfection reagent |
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