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 0.15 µg 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%
60% 55% 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 - UPR2580, Montpellier) for nicely providing 600 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 .
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