HIGHLIGHT:

  • Suitable for transfection of DNA, mRNA, siRNA, and miRNA, as well as for cotransfection of DNA and RNA
  • Suitable for stable and transient transfection
  • Suitable for adherent and suspended cells.
  • Minimized toxic effects
  • Optimized transfection rates
  • Free of serum inhibition
  • Complete library of user reports
  • Wide range of references in specialized literature

METAFECTENE® PRO is a liposome-based transfection reagent consisting of a mixture of polycationic and neutral lipids. It is an advanced product based on METAFECTENE. The structural changes of the cationic lipids that comprise them result in greater efficacy and less toxicity.

METAFECTENE® PRO is part of a temporary development series of Biontex transfection reagents, which includes, in particular, the following reagents: METAFECTENE® <METAFECTENE® PRO <K2® Transfection System <K4® Transfection System. With the degree of development the probability of a successful transfection increases (see also General Selection Aid).

Low cytotoxicity and lack of serum inhibition allow a wide range of applications. METAFECTENE® PRO is suitable for the transient and stable transfection of mammalian cell lines and primary cells with DNA (plasmids, bacmids), RNA (mRNA, miRNA, and siRNA), and modified nucleic acids (antisense oligonucleotides). Areas of application are particularly the production of proteins, antibodies, and viruses (eg adenovirus, AAV, lentivirus), cotransfections of different nucleic acids, genome editing (eg CRISPR / Cas9, CRISPR / Cpf1), and gene silencing. (gene deletion).

Transfection methods

Various methods are used to introduce genetic material into eukaryotic cells:

Viral transfection uses genetically modified viruses that are no longer pathogenic and carry the gene to be introduced. In this method, problems include stable, often non-directional, integration of the gene into the host cell genome, laborious production of viruses, and strong immune response. Physical methods such as electroporation or microinjection are expensive and of limited use in routine procedures.

Different transfection methods | Biontex

Chemical methods include calcium phosphate precipitation, transfection with cationic polymers, and, among the most effective methods, lipofection with cationic lipids. Biontex transfection reagents (DOTAP, METAFECTENE®, METAFECTENE® PRO, METAFECTENE® SI, K2® Transfection System, K4® Transfection System) belong to the latter group.

Cationic lipid transfection

In aqueous solutions, cationic lipids form vesicles with a bilayer lipid sheet, known as liposomes. When liposomes encounter nucleic acids, they reform into lipid complexes of nucleic acids called lipoplexes that can be actively taken up by eukaryotic cells through endocytosis. In this case, the lipoplex enters the cell cytosol through the endosomes.

The endosomal structure is destroyed by increasing the osmotic pressure created by the buffering action of the lipids within the endosomes and by the fusion of the lipid with the endosomal membrane. The ability of lipid to destroy endosomes is one of the main characteristics of an efficient transfection reagent.

DNA transfection

DNA that enters the cell cytosol cannot penetrate the membrane of the cell nucleus (“nuclear barrier”). Therefore, access to the nucleus is only possible if the nuclear membrane dissolves during mitosis. For this reason, the rate of cell division is critical in DNA transfection and must be as high as possible for efficient transfection.

RNA transfection (siRNA / miRNA / mRNA)

For RNA transfection, there is no “nuclear barrier”, since RNA does not need to enter the nucleus to develop its biological effects. As a consequence, the rate of cell division has no influence. Lipofection is a complex process in which the amount of lipoplex and the ratio of genetic material in relation to the transfection reagent, that is, the composition of the lipoplex must be precisely optimized for each cell type.

Transfection reagents and methods show different performance profiles depending on the type of nucleic acid and the type of cell used. Without reference, it is not possible to predict the suitability of a reagent or method with respect to a special application and must be determined empirically. Our “Transfection Reagent Selection Guide” helps you select a suitable transfection reagent for your application. We also offer free samples, so you can test whether one of our transfection reagents is suitable for your purposes, at no cost.