Abstract
This paper describes a method by which molecules that are impermeable to cells are encapsulated in dye-sensitized lipid nanocapsules for delivery into cells via endocytosis. Once inside the cells, the molecules are released from the lipid nanocapsules into the cytoplasm with a single nanosecond pulse from a laser in the far red (645 nm). We demonstrate this method with the intracellular release of the second messenger IP(3) in CHO-M1 cells and report that calcium responses from the cells changed from a sustained increase to a transient spike when the average number of IP(3) released is decreased below 50 molecules per nanocapsule. We also demonstrate the delivery of a 23 kDa O(6)-alkylguanine-DNA alkyltransferase (AGT) fusion protein into Ba/F3 cells to inhibit a key player BCR-ABL in the apoptotic pathway. We show that an average of ∼8 molecules of the inhibitor is sufficient to induce apoptosis in the majority of Ba/F3 cells.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Animals
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Biological Transport
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CHO Cells
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Calcium / metabolism
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Cricetinae
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Cricetulus
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Enzyme Inhibitors / chemistry
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Enzyme Inhibitors / pharmacology
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Fusion Proteins, bcr-abl / antagonists & inhibitors
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Fusion Proteins, bcr-abl / metabolism
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HEK293 Cells
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Humans
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Inositol 1,4,5-Trisphosphate / chemistry
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Inositol 1,4,5-Trisphosphate / metabolism
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Inositol 1,4,5-Trisphosphate Receptors / metabolism
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Intracellular Space / metabolism*
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Intracellular Space / radiation effects*
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Lasers
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Light*
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Lipids / chemistry
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Nanocapsules* / chemistry
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O(6)-Methylguanine-DNA Methyltransferase / antagonists & inhibitors
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O(6)-Methylguanine-DNA Methyltransferase / metabolism
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Photolysis
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Time Factors
Substances
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Enzyme Inhibitors
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Inositol 1,4,5-Trisphosphate Receptors
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Lipids
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Nanocapsules
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Inositol 1,4,5-Trisphosphate
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O(6)-Methylguanine-DNA Methyltransferase
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Fusion Proteins, bcr-abl
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Calcium