Abstract
Intracellular delivery of biologically active proteins remains a formidable challenge in biomedical research. Here we show that biomedically relevant enzymes can be delivered into cells using a new DNA transfection reagent, lipofectamine 3000, allowing assessment of their intracellular functions. We also show that the J774.2 macrophage cell line exhibits unusual intracellular uptake of structurally and functionally distinct enzymes providing a convenient, reagent-free approach for evaluation of intracellular activities of enzymes.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amiloride / pharmacology
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Cell Count
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Cell Line
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Cell Survival / drug effects
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Dextrans / chemistry
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Epithelial Cells / cytology
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Epithelial Cells / drug effects
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Epithelial Cells / metabolism*
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Fluorescein-5-isothiocyanate / analogs & derivatives
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Fluorescein-5-isothiocyanate / chemistry
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Gene Expression
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Genes, Reporter
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Humans
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Liposomes / chemistry
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Liposomes / pharmacology*
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Luminescent Proteins / genetics
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Luminescent Proteins / metabolism
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Macrophages / cytology
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Macrophages / drug effects
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Macrophages / metabolism*
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Neurons / cytology
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Neurons / drug effects
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Neurons / metabolism*
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Organ Specificity
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Protein Synthesis Inhibitors / chemistry
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Protein Synthesis Inhibitors / pharmacology
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Red Fluorescent Protein
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Ribosome Inactivating Proteins, Type 1 / chemistry
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Ribosome Inactivating Proteins, Type 1 / pharmacology
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Saporins
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Transfection / methods*
Substances
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Dextrans
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Liposomes
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Luminescent Proteins
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Protein Synthesis Inhibitors
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Ribosome Inactivating Proteins, Type 1
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fluorescein isothiocyanate dextran
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Amiloride
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Saporins
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Fluorescein-5-isothiocyanate