Magnetofection TM the new gene transfection …

1 MagnetofectionTMthe newgene transfection technologyMagnetofectionTMis a novel, simple and highly efficientmethod to transfect cells in culture. Table of Magnetofection - Optimization Protocol in 96-well High Throughput Magnetofection - Viruses CombiMAG Magnetofection - siRNA PolyMAG siRNA CombiMAG Magnetofection - Suspension CombiMAG Example Protocol for Fugene (Roche) CombiMAG Example Protocols for other Cationic Lipid Magnetofection in 6-, 12-, 24- or 96-well Plate Formats, T-75 Culture General General Kits Contents12 Avalaible MagnetoFACTOR plates11 Utensils for Magnetofection Nucleic Acids Dose Response and transfection Magnetofection Description2 TechnologyTechnologyMagnetofection is a novel, simple and highly efficient method to transfect cells in culture.

2 It exploits magnetic force exerted upon gene vectors associated with magnetic particles to draw the vectors towards, possibly even into, the target cells. In this manner, the full vector dose applied gets concentrated on the cells within a few minutes so that 100% of the cells get in contact with a significant vector dose. This has several important consequences: Greatly improved transfection rates in terms of percentage of cells transfectedcompared to standard transfection .

3 Up to several thousand fold increased levels of transgene expression compared to standard transfections upon short-term incubation. High transfection rates and transgene expression levels are achievable with extremely low vector doses, which allows to save expensive transfection reagents. Extremely short process time. A few minutes of incubation of cells with gene vectors are sufficient to generate high transfection efficiency, compared to several hours with standard procedures.

4 Chemicell GmbH Magnetofection Description3As the manufacturer of the Magnetofection technology, chemicelloffers two types of ready-to-use Magnetofection a universally applicable magnetic particle preparation for high efficiency nucleic acid delivery. It is mixed in a one-step procedure with the nucleic acid to be transfected and has been used successfully with plasmid DNA, antisense oligonucleotides and siRNAs. CombiMAGis a magnetic particle preparation designed to be combined with any commercially available transfection reagent such as polycations and lipids and can be associated with plasmid DNA, antisense oligonucleotides, siRNAs or viruses.

5 It allows you to create your own magnetic gene vector based on your favouritetransfection (expression level) 050100150200250process time (min)050100150200response (expression level) ( g DNA)paramagneticvehicle no magnetic fieldstandardgene transferparamagneticvehicle plusmagnetic fieldDose response profile in NIH 3T3 cells using Lipofectamine (Invitrogen) CombiMAG with and without positioning on the MagnetoFACTOR plate for 15 min. Luciferase expression was assayed after 24 kinetics: NIH 3T3 cells were incubated with GenePorter ( gene Therapy Systems) CombiMAG with and without positioning on the Magneto-FACTOR plate for the indicated time spans.

6 Luciferase expression was assayed after 24 NotificationThe Magnetofection reagents and all of its components are developed, designed, intended and soldfor research use only. They are not to be used for human diagnostic or any drug are registered Magnetofection Nuclec Acids Dose Response and transfection Kineticschemicell GmbH Magnetofection is generally applicable for adherent cells and has been tested witha variety of immortalized cell lines and primary cells listed below. If a particular celltype or cell line is not listed this does not mean that Magnetofection would not work.

7 Also for the cells listed, some reagents have not been tested so far, as indicated by (not determined). TheCombiMAG reagent can be combined with any polycationic and lipidictransfection reagent, and also with adenoviral and retroviral vectors. In some cases, references are made in the footnotes to very successful combinations withcommercially available reagents that have been tested so human umbilical vein endothelial cells positioned on theMagnetoFACTOR plate were incubated for 15 min with a Cy3 fluorescence-labeled antisense-oligonucleotide complexed with Effectene (Qiagen; left) or Effectene + CombiMAG (right).

8 (Data kindly provided by F. Kroetz. Ludwig-Maximilians University Munich) Application siRNA Antisense Oligonucleotides Plasmid DNAC ombiMAGPolyMAGType of nucleic acid / viruschemicell GmbH Magnetofection , ,++humanalveolar basal ,++ ,++ ,+ ,+ ,+ ,+ ,+ ,+ ,+ , ,+ ++humanovarianCHOA dherent Cells++++ ++ +++++CombiMAG* adenocarcinomaHCT 116-+humanbreast , , , ,+humanbreast +humanbladder ,+humanairway , , , , , ,+mousefibroblastsNIH , , , ,+ , , ,+humankidneyHEK.

9 +humancervix carcinomaHeLaRef. lineCell lineTechnology5chemicell GmbH Magnetofection + +humanfibrosarcomaHT-1080-++humanforeski n , ,++humanhepatocellular carcinomaHep G2-+ ,++humankidney293 ,+ +humanlung carcinomaSK-MES-1-++humanlung, embryonicMRC 5-+ ,+ glioblastomaNYGM-+ ++humanlung + + ,+ ,+ lymphoblastic , +humanmicrovascular , , + + , neck , +humantransformed , ,+ , + cell lung carcinomaA549-+ sarcomaSaOS-2-+ sarcomaBTK-143-+ +humanlung epithelial cellsBEAS-2B+++ gland.

10 GmbH Magnetofection + cell ++humanlung , , +humanmucoepidermoid +humansmall cell lung + +humanbreast adenocarcinomaMDAMB231-+ bladder + + , +mousecolon + mammary +mouseembryonal +mousefibroblast, connective , + + smooth + lymph +mousemurine + marrow-derived ++mouseenteroendocrine I ++mouseB-cell linebTC-tet+ ++ ++ +ratdaughter cell line of PC12PC6-3-+ratadrenal , + , + (cl2) neuroblastHT-22 Technology7chemicell GmbH Magnetofection + +ratskeletal muscle , , ,+ , neck +ratintestinalRIE1- + ++ +monkeykidneyCV-1-+ , , + + (UV) resistant +hamsterlung fibroblastsV79-+ +caninekidneyMDCK-+ + + Cells (epicard)EMC-+ +fishbone (chondrocyte-like)VSa13 Suspension Cells+++ ++CombiMAG*-+humanhistiocytic myeloid lymphomaK562-+humanB-cell , T-cell myeloid leukemiaTHP-1 Ref.