Viromer® in metabolic research

Human obesity is one of the most important diseases of the western world and a leading cause of death worldwide. It increases the likelihood for the manifestation of cardiovascular diseases, type 2 diabetes, different kinds of cancer, depression and asthma. Since obesity has been found to be one of the most important public health problems of the 21st century, research started to focus more and more on the metabolic system and the defective consequences for the human body. What is the cause of obesity? How can we prevent it? Which cellular mechanisms are involved? Can we treat the cells with medication to minimize or stop the production of fatty acids. What ways do we have to force these cells to lose stored fatty acids?

The morbidity due to the increased number of fat cells includes diabetes, cancer, cardiovascular disease and non-alcohol associated liver diseases. The in vitro transfection of these related cells is an unevitable necessity in research to understand the molecular mechanisms behind, on the way to address the fight against obesity related diseases. Since Viromer® transfection reagents are polymer based and lipid-free, there is no interaction with the cell metabolism and in particular with the lipid metabolism, which makes them the perfect transfection reagent for adipocytes and liver cells in comparison to lipid based reagents. The Viromer® reagents provide excellent results in adipocytes, hepatocytes, fibroblasts, kidney cells and pancreatic beta cells – as well as for knockdown studies with siRNA or overexpression experiments with pDNA or mRNA.

Expression of GFP in rat hepatocyte cell line FAO transfected with Viromer® YELLOW.

“In comparison to 3 other transfection reagents Viromer YELLOW was most sensitive with highest transfection efficiency 48h post transfection.” C. Klingler, University Hospital Tübingen, Germany

siRNA-mediated gene silencing

Up to 80-100% complete specific gene knock-downs have been achieved by transfecting siRNA into different kinds of adipocytes, hepatocytes, fibroblasts or kidney cells using Viromer® BLUE  or  Viromer® GREEN:

Knock down of gene X expression in pre-adipocytes and mature adipocytes with Viromer® BLUE. Use of NIH/3T3-L1 cells. → max. knockdown after 72h

Knock-down of ppar-alpha gene expression in primary hepatocytes after siRNA transfection with Viromer® BLUE (cells freshly isolated from mouse liver, transfection after 24h, test with 10nM and 100nM siRNA → 100nM yielded a complete knock-down)

Protein Expression

Microscopic observations of MEFs (mouse embryonic fibroblasts) transfected with Drp1-YFP, co-stained with mitotracker red and DAPI, by using Viromer® YELLOW.

  • Overnight transfection

  • 6-well plate format, standard protocol

Expression of a glutamate membrane transporter tagged with GFP in COS-7 fibroblasts transfected via a plasmid vector with Viromer® RED.

  • approx. 80% of transfected cells

Colors

Viromer® BLUE | Viromer® GREEN | Viromer® RED | Viromer® YELLOW

transfection efficiency

< 30% | 30-50% | 50-80% | > 80%

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Type siRNA mRNA pDNA Data
A-375       
AD-293     
C3H/10T1/2      See data
COS-7      See data
FAO      See data
Fibroblasts, primary human + murine               See data Publications
H295R         See data
HaCaT            See data
Hep G2               Publication
HepaRG      See data
Hepatocytes, primary mouse          See data Publication
HUH-7        
INS-1    
Keratinocytes, primary human + mouse     See data Publications
Kidney cell line, self-established     
L929          See data
LLC-PK1      See data
MDCK                  See data
MDOK   
MEFs    See data Publication