Intracellular delivery of bioactive molecules using light-addressable nanocapsules.
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.
Chemistry Department, University of Washington, Seattle, Washington, USA.
Kimberly A D
KA
Gregersen
Zachary B
ZB
Hill
Jennifer C
JC
Gadd
Bryant S
BS
Fujimoto
Dustin J
DJ
Maly
Daniel T
DT
Chiu
D000818:Animals; D001692:Biological Transport; D016466:CHO Cells; D002118:Calcium / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology; D016044:Fusion Proteins, bcr-abl / Q000037:antagonists & inhibitors / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D015544:Inositol 1,4,5-Trisphosphate / Q000737:chemistry / Q000378:metabolism; D053496:Inositol 1,4,5-Trisphosphate Receptors / Q000378:metabolism; D042541:Intracellular Space / Q000378:metabolism* / Q000528:radiation effects*; D007834:Lasers; D008027:Light; D008055:Lipids / Q000737:chemistry; D053769:Nanocapsules / Q000737:chemistry; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000037:antagonists & inhibitors / Q000378:metabolism; D010782:Photolysis; D013997:Time Factors
D004791:Enzyme Inhibitors; D053496:Inositol 1,4,5-Trisphosphate Receptors; D008055:Lipids; D053769:Nanocapsules; D015544:Inositol 1,4,5-Trisphosphate; D019853:O(6)-Methylguanine-DNA Methyltransferase; D016044:Fusion Proteins, bcr-abl; D002118:Calcium