Cationic lipid nanodisks as an siRNA delivery vehicle.
The term nanodisk (ND) describes reconstituted high-density lipoprotein particles that contain one or more exogenous bioactive agents. In the present study, ND were assembled from apolipoprotein A-I, the zwitterionic glycerophospholipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), and the synthetic cationic lipid 1,2-dimyristoyl-3-trimethylammonium-propane (DMTAP). ND formulated at a DMPC:DMTAP ratio of 70:30 (by weight) were soluble in aqueous media. The particles generated were polydisperse, with diameters ranging from ∼20 to <50 nm. In nucleic acid binding studies, agarose gel retardation assays revealed that a synthetic 23-mer double-stranded oligonucleotide (dsOligo) bound to DMTAP containing ND but not to ND formulated with DMPC alone. Sucrose density gradient ultracentrifugation studies provided additional evidence for stable dsOligo binding to DMTAP-ND. Incubation of cultured hepatoma cells with DMTAP-ND complexed with a siRNA directed against glyceraldehyde 3-phosphate dehydrogenase showed 60% knockdown efficiency. Thus, incorporation of synthetic cationic lipid (i.e., DMTAP) to ND confers an ability to bind siRNA and the resulting complexes possess target gene knockdown activity in a cultured cell model.
a Children's Hospital Oakland Research Institute, 5700 Martin Luther King Jr. Way, Oakland, CA 94609, USA.
Mistuni
M
Ghosh
Gang
G
Ren
Jens B
JB
Simonsen
Robert O
RO
Ryan
D002412:Cations / Q000737:chemistry; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry*; D004337:Drug Carriers / Q000737:chemistry*; D016503:Drug Delivery Systems; D049329:Nanostructures / Q000737:chemistry*; D010316:Particle Size; D034741:RNA, Small Interfering / Q000737:chemistry / Q000378:metabolism*; D013499:Surface Properties
D002412:Cations; D004337:Drug Carriers; D034741:RNA, Small Interfering; D004134:Dimyristoylphosphatidylcholine
1;2-dimyristoyl-3-trimethylammonium-propane; apolipoprotein; apolipoprotéine; cationic lipid; cellules HepG2; delivery and HepG2 cells; dimyristoyltriméthylaminopropane; distribution; lipide cationique; nanodisk; nanodisque; pARNi; siRNA