Magnetic and acoustically active microbubbles loaded with nucleic acids for gene delivery.
Targeted gene or drug delivery aims to locally accumulate the active agent and achieve the maximum local therapeutic effect at the target-site while reducing unwanted effects at nontarget sites. A further development of the magnetic drug-targeting concept is combining it with an ultrasound-triggered delivery using magnetic microbubbles as a carrier for gene or drug delivery. For this purpose, selected magnetic nanoparticles (MNPs), phospholipids, and nucleic acid are assembled in the presence of perfluorocarbon gas into flexible formulations of magnetic lipospheres or microbubbles. This manuscript describes the protocols for preparation of magnetic lipospheres and microbubbles for nucleic acid delivery, and it also describes the procedures for labeling the components of the bubbles (lipids, MNPs, and nucleic acids) for the visualization of the vectors and their characterization, such as magnetic responsiveness and ultrasound contrast effects. Protocols are given for the transfection procedure in adherent cells, evaluation of the association of the magnetic vectors with the cells, reporter gene expression analysis, and cell viability assessment.
Institute of Experimental Oncology and Therapy Research, Klinikum rechts der Isar, Technische Universität München, Munich, Germany. dialekti.vlaskou@lrz.tu-muenchen.de
Dialechti
D
Vlaskou
Christian
C
Plank
Olga
O
Mykhaylyk
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D060328:Magnetic Phenomena; D051379:Mice; D045423:Microbubbles; D041681:NIH 3T3 Cells; D053758:Nanoparticles / Q000737:chemistry / Q000633:toxicity; D010957:Plasmids / Q000235:genetics; D011094:Polyethyleneimine / Q000737:chemistry; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D014162:Transfection / Q000295:instrumentation*; D014465:Ultrasonics / Q000295:instrumentation*
D005456:Fluorescent Dyes; D034741:RNA, Small Interfering; D011094:Polyethyleneimine; D004247:DNA