- transfections of cell lines and primary cells with nucleic acids (plasmid DNA, siRNA, oligonucleotides)
- for adherent and suspension cells
- expression and analysis of genes
- suited also for automated high-thoughput transfections
- very high transfection efficiencies with a wide range of cell types (many common cell lines as well as primary cells)
- low cytotoxicity with most commonly used cell lines
- up to several thousand fold increased levels of transgene expression after short-term incubation with the transfection reagent
- successfully used with many hard-to-transfect cells such as primary human cells
- easy, fast and reproducible protocol with different assay formats
- functional with serum and serum-free
- highly cost-effective - no need for expensive instruments (like electroporator or particle gun)
- saves time and material (allows to apply lower amounts of nucleic acid for transfection!)
MATra-A Reagent consists of a suspension containing specifically coated magnetic nanoparticles (MagTag™) which can be loaded with the nucleic acid of interest (e.g. plasmid DNA, siRNA or oligonucleotides). Exploiting magnetic force (using a strong magnet plate that is placed beneath the target cells to be transfected), the complex consisting of the magnetic nanoparticles and the bound nucleic acids is rapidly drawn towards the cells where it is deposited at a high concentration directly on the outer cell membrane resulting in a very fast and efficient transfection of the cells. The uptake of this complex by the host cell is carried out via normal endocytosis. The transfection rate can thus be clearly increased - also with difficult to transfect cells such as primary cells - and the amount of nucleic acid required for successful transfection can be significantly reduced which in turn further reduces cytotoxicity. MATra-A Reagent can be used for the transfection of adherent and suspension cells; for transfection of suspension cells see also MATra-S Immobilizer.
MATra-A Reagent has been successfully used to deliver siRNA into many different cell lines, such as carcinoma cell lines (see Examples & Applications). However, for optimized siRNA transfection and prolonged gene silencing PromoKine provides the specifically developed MATra-siRNA Reagent.
siRNAs are small interfering/inhibitory sequence-specific RNA molecules that are able bind to a complementary sequence of a target mRNA, block translation of that target mRNA, and initiate its degradation. Consequently, synthesis of the corresponding protein of interest is prevented and the specific gene function is knocked-out. This process is termed RNA interference or gene-silencing and it is used to elucidate gene functions. Specifically designed small inhibitory RNA oligonucleotides (siRNAs) allow transient inhibition of gene expression in vitro and in vivo.
MATra-siRNA contains specifically coated magnetic nanoparticles, which are optimized to effectively bind and stabilize siRNA. It provides optimal delivery of siRNA into many different cell types enabling efficient and prolonged gene silencing. It can also be used to deliver mRNA, miRNA and oligonucleotides.
Suspension cells have to be made "quasi-adherent" first by incubating them with the MATra-S Immobilizer reagent which contains magnetic nanoparticles (MagTag™) that are however not identical with the MATra-A Reagent particles (which only bind nucleic acids). The specifically coated MATra-S Immobilizer particles bind the floating suspension cells. By applying a strong magnetic force (placing a strong magnet plate beneath the respective cell culture flask or multi-well plate), the suspension cells are drawn to the bottom of the cell culture vessel lying there quasi-adherent and ready for the following actual transfection procedure for which either MATra-A Reagent or MA Lipofection Enhancer in combination with a transfection reagent of choice can be used.
MA Lipofection Enhancer*:
Transfection with common lipidic (liposomal) lipofection reagents (e.g. Lipofectamine, Lipofectin, TransFast, ClonFectin, TransFectin or GenePorter) or polycationic (non-liposomal) transfection reagents such as PromoFectin, FuGene, TransIT-LT, PolyFect/SuperFect or GeneJammer can be significantly enhanced by magnetic assistance ("Magnet Assisted Lipofection"). In this case, the nucleic acid of interest has to be combined with MA Lipofection Enhancer in the presence of a common transfection reagent. The formulation of MA Lipofection Enhancer, a suspension containing coated magnetic nanoparticles (MagTag™), has been optimized for use with common lipofection reagents but does also work with other non-liposomal, cationic transfection reagents. The MA Lipofection Enhancer binds the transfection reagent of choice that in turn binds the nucleic acid of interest. This complex - consisting of MA Lipofection Enhancer, the transfection reagent of choice and the respective target nucleic acid (e.g. plasmid DNA or siRNA) - is then drawn rapidly towards the cell, deposited directly on the outer cell membrane and eventually internalized by endocytosis.
* The MA Lipofection Enhancer is not essential for many cells. For critical cell lines we recommend first trying MATra-A Reagent. If you want to benefit from the fast and efficient Magnet-assisted Transfection Technology and want to retain your traditional lipofection reagent you may use MA Lipofection Enhancer with PromoFectin or any other transfection reagent of your choice in parallel to find the optimal solution for your cells of interest!