PolyMag Result s Magnetofection™ is a novel, simple and highly efficient method to transfect cells in culture and in vivo. It exploits magnetic force exerted upon gene vectors associated with magnetic particles to drive the vectors towards, possibly even into, the target cells. In this manner, the complete applied vector dose 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: 1. Greatly improved transfection rates in terms of percentage of cells transfected compared to standard transfections. 2. Up to several thousand folds increased levels of transgene expression compared to standard transfections. 3. High transfection rates and transgene expression levels are achievable with extremely low vector doses, which allow saving expensive transfection reagents. 4. Extremely short process time in comparison to standard procedures. A few minutes of incubation of cells with gene vectors are sufficient to generate high transfection efficiency. OZ Biosciences offers three types of ready-to-use reagents: SilenceMag specific to siRNA delivery, CombiMag designed to be associated with all vectors: transfection reagents and viruses and PolyMag . PolyMag has been designed and used successfully to deliver all types of nucleic acids such as plasmid DNA, RNA, Oligonucleotides & siRNA. PolyMag is a universally applicable magnetic particle preparation for high transfection efficiency. Nucleic acids to be transfected and the magnetic particles are mixed in a one-step procedure. Nucleic Acid Types Nucleic Acid or Virus Type PolyMag SilenceMag CombiMag DNA (plasmid) √ NA √ Oligonucleotides √ ND √ mRNA √ ND √ siRNA √ √ √ dsRNA ND √ √ shRNA ND √ √ Adenovirus NA NA √ Retro virus NA NA √ Cell Types PolyMag is generally applicable on numerous cell types. This technology has been tested successfully on a variety of immortalized and primary cells. Please consult our updated list of cells successfully tested available on the website: www.ozbiosciences.com . If a particular cell type or cell line is not listed, this does not imply that PolyMag is not going to work. OZ Biosciences is going to frequently adjust this list. OZ Biosciences – PolyMag Results Page 1 of 5 Transfec tion of Primary Cells Standard Transfection Magnetofection Confluent Primary Human Keratinocytes Transfected with a commercial reagent L or PolyMag Reporter Gene: GFP We are grateful to the laboratories of Dr. C. Plank (Technical University, Munich) for kindly providing these data. 50 Primary Porcine Aortic Endothelial Cells PolyMagMagnetofecti Transfected with a commercial reagent P or 40 Reagent(PolyMa P PolyMag Standard Reporter Gene: GFP (PEI) Culture dish: 96-well plate 30 We are thankful to Dr. F. Kroetz (Ludwig- 20 Maximilians University, Munich) for nicely providing these data. 10 0 DNA dose (ng) Transfection of Cell Lines Standard Transfection Magnetofection CT -26 Colon Carcinoma / Mouse Transfected with a commercial reagent D or PolyMag DNA Reporter Gene: GFP µ 0.15 g Culture dish: 24-well plate We are grateful to the laboratories of Dr. C. Plank (Technical University, Munich) for kindly providing these data. DNA 0.3 µg OZ Biosciences – PolyMag Results Page 2 of 5 Various examples of PolyMag transfection efficiency (24-well plate; 0.5 l PolyMag & 0.5g DNA/ well): COS 7 N2A Hep2 HeLa Monkey Kidney Fibroblast Mouse Neuroblastoma Human Epithelial Human Epithelial Carcinoma PolyMag Transfection Efficiency 100% 90% 85% 80% 90% 80% 75% 75% 70% 70% 55% 60% 50% 50% 50% 40% 30% transfection% efficiency 20% 10% 0% 293 BHK-21 CHO COS7 CT-26 HaCaT HT1080 MCF-7 N2A Cells Improved and Optimized PolyMag Formulation OZ Biosciences has developed a new very efficient PolyMag formulation for Magnetofection TM . This improved PolyMag II reagent gives reliable higher transfection efficiencies in numerous cell types (see figures), transports all type of nucleic acids (DNA, siRNA, ODN). This PolyMag optimized formulation is the actual commercialized PolyMag reagent. Most of the herein described results were obtained with the new optimized PolyMag formulation. Transfection efficiency PolyMag - HaCaT cells 1200 1000 PolyMag I PolyMag II 800 We are thankful to Dr. L. Journot (CNRS - 600 UPR2580, Montpellier) for nicely providing these data. 400 200 Luciferase(count/s) activity 0 2 4 8 µg pRSV -Luc/plate OZ Biosciences – PolyMag Results Page 3 of 5 Delivery of Oligonucleotides Standard Transfection Magnetofection HUVEC -C (Primary) / Human Transfected with a commercial reagent E or PolyMag Fluorescent Oligonucleotides Culture dish: 24-well plate ODN: 200 ng / well We are grateful to Dr. F. Kroetz (Ludwig- Maximilians University, Munich) for kindly providing these data. siRNA Delivery Efficiency GFP stably transfected HeLa cells . Target siRNA : GFP Cell culture dish : 96-well plate Transfection volume : 200 l. Control PolyMag + siRNA/10nM PolyMag + siRNA/40nM Primary Human Umbilical Vein Endothelial Ce lls TF # 1 Gene Silencing mediated by PolyMag/siRNA (HUVEC). Target siRNA : Transcription factor # 1 100 90 Control siRNA : Transcription factor # 2 (Belongs to TF#1 family) 80 70 Control siRNA : GFP 60 50 Cell culture dish : 60 mm 40 Normalized 30 SiRNA final concentration: 25nM (2x treatments) expression unit 20 10 We are grateful to Dr. D. Mathieu (CNRS- 0 GFP TF#1 TF#2 TF #1 Control UMR5535, Montpellier) for kindly providing these + # 2 data . siRNA OZ Biosciences – PolyMag Results Page 4 of 5 Cytotoxicity Electroporation vs. Magnetofection 100 90 80 70 60 50 transfection - 40 % of% cells living 30 post 20 10 0 Electroporation PolyMag Bibliographic References Please consult our list of references available on the website: www.ozbiosciences.com . OZ Biosciences – PolyMag Results Page 5 of 5 .