Tuesday, June 29, 2010

Using MATRa for siRNA Transfection of Carcinoma Cell Lines

MATRaTM -Magnet Assisted Transfection is an easy-to-handle, very fast and highly efficient technology to transfect cells in culture with siRNA. Multiple successes with the system includes Carcinoma Cell Lines.

Efficient transient transfection of siRNA in head and neck cancer cells. The cell line ANT-1 was transiently transfected with MATra-A (1 µl/1 µg DNA) in a 6 well format (5 x 105 cells/cavity) with siRNA against protein 1 (100 nM). After 24 hours total RNA was isolated and expression of protein 1-specific mRNA determined by RT-PCR (upper lane). SiRNA 13 are three different oligonucleotide sequences. Control for consistent loading and cDNA quality: expression of ubiquitary GAPDH mRNA (lower lane).
Protein 2 expression in head and neck cancer cells GHD-1. GHD-1 cells (5 x 105 cells/cavity of a 6 well plate) were transiently transfected with two different siRNAs against protein 2. Expression of protein 2 was detected with specific antibodies in an immunoblot 72 hours after transfection with MATra-A (1 µl / 1 µg DNA). As control ubiquitary β-actin was detected as well.
Treating the carcinoma cells with specific siRNA caused a clear inhibition of protein 1/protein 2 expression which indicates high transfection efficiencies.
(Data kindly provided by Rauch, Schaffrik, Ahlemann and Gires, LMU Munich and GSF, Munich, Germany).

 "After having tested MATra in a variety of experimental set ups we can summarize the following advantages:

  • High transfection efficiency 
  •  Easier to handle  
  • High reproducibility
  • Serum compatibility
  • Low sensibility against cell confluence"

Dr. Oliver Gires, LMU Munich, Germany

Friday, June 25, 2010

siRNA and i-Fect for the Study of Retinal Disease

We continue to add new references to the many ways researchers are using i-FectTM to increase the potency of siRNA delivery.

This is a new reference from the book entitled Retinal Degenerative Diseases: Laboratory and Therapeutic Investigations By Robert E. Anderson, Joe G. Hollyfield, Matthew M. Lavail.

In this study, researchers used i-Fect to transfect and immortalized cell line from Mouse cones (661W) expressing ELOVL4. Using siRNA designed to silence the ELOVL4 gene, they used i-Fect + siRNA to transfect cells cultured at a density of 2X105. Knockdown was achieved as confirmed by western blot analysis.