Liposomal magnetofection.
In a magnetofection procedure, self-assembling complexes of enhancers like cationic lipids with plasmid DNA or small interfering RNA (siRNA) are associated with magnetic nanoparticles and are then concentrated at the surface of cultured cells by applying a permanent inhomogeneous magnetic field. This process results in a considerable improvement in transfection efficiency compared to transfection carried out with nonmagnetic gene vectors. This article describes how to synthesize magnetic nanoparticles suitable for nucleic acid delivery by liposomal magnetofection and how to test the plasmid DNA and siRNA association with the magnetic components of the transfection complex. Protocols are provided for preparing magnetic lipoplexes, performing magnetofection in adherent and suspension cells, estimating the association/internalization of vectors with cells, performing reporter gene analysis, and assessing cell viability. The methods described here can be used to screen magnetic nanoparticles and formulations for the delivery of nucleic acids by liposomal magnetofection in any cell type.
Mykhaylyk
Olga
O
Institute of Experimental Oncology and Therapy Research, Technische Universität München, München, Germany.
Sánchez-Antequera
Yolanda
Y
Vlaskou
Dialekti
D
Hammerschmid
Edelburga
E
Anton
Martina
M
Zelphati
Olivier
O
Plank
Christian
C
D045744:Cell Line, Tumor; D002470:Cell Survival; D004247:DNA / Q000008:administration & dosage*; D015870:Gene Expression; D017930:Genes, Reporter; D006801:Humans; D007501:Iron / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry*; D008280:Magnetics; D053758:Nanoparticles / Q000737:chemistry*; D010957:Plasmids / Q000008:administration & dosage; D034741:RNA, Small Interfering / Q000008:administration & dosage*; D014162:Transfection
D008081:Liposomes; D034741:RNA, Small Interfering; D004247:DNA; D007501:Iron