In cell biology, transfection is the term given to the introduction of foreign genetic material into eukaryotic cells. In addition, the distinction is made between temporary introduction into the host cell transient transfection) and permanent integration into the genome (stable transfection). The process of transfection basically corresponds to that of bacterial transformation, albeit under a different name.
A variety of methods are used to introduce genetic material into eukaryotic cells: Viral transfection uses genetically modified viruses which are no longer pathogenic and carry the gene to be introduced. In this method problems include the frequently non-directional stable integration of the gene into the genome of the host cell, the laborintensive production of the viruses and the strong immunological response.
Physical methods such as electroporation or microinjection are cost-intensive and of limited use in routine procedures. Chemical methods include calcium phosphate precipitation, transfection using cationic polymers and - among the most efficient methods – lipofection. Our areas of core competency are in the field of lipofection using cationic lipids. Our products provide users with the optimum technology for their transfection assays in each case.
In aqueous solutions, cationic lipids form vesicles with a bilayer lipid sheet, known as liposomes. When liposomes encounter nucleic acids they re-form into nucleic acid lipid complexes called lipoplexes which can be actively taken up by eukaryotic cells by means of endocytosis. In this case, the lipoplex enters into the cell cytosol via the endosomes1.
The endosomal structure is destroyed by increasing the osmotic pressure created by the lipids' buffering action within the endosomes and by the fusion of the lipid with the endosomal membrane. The ability of a lipid to destroy endosomes is one of the main characteristics of a transfection reagent. Our METAFECTENE® transfection products based on the core product METAFECTENE® (cat. no. T020) feature structural elements designed to optimally promote precisely these properties (RMA technology).
DNA which is introduced into the cell cytosol cannot penetrate the membrane of the cell nucleus ("nuclear barrier"). Access to the nucleus is thus only possible if the nuclear membrane dissolves during mitosis. For this reason, the cell division rate is critical in DNA transfection and must be as high as possible for efficient transfection.
Another key criterion for successful transfection is the toxicity of the reagent – and thus the condition of the cells. We have set new standards in this area by introducing TOP technology as a constituent of our transfection reagents since our development of METAFECTENE® PRO (cat. no. T040).
Transfection is a complex process in which the lipoplex volume or proportion of genetic material in relation to transfection reagent must be precisely optimized for each cell type. The numerous optimization stages required to achieve satisfactory levels of transfection efficiency are extremely timeconsuming and require large volumes of reagents and other substances – a not inconsiderable cost factor. Now Biontex has developed FEE technology and incorporated it into METAFECTENE® EASY (cat.no. T090), the first third-generation transfection reagent. This reagent is used in a single predefined quantity, thus saving time, effort and costs while delivering excellent transfection results.
Whether users seek a reagent to maximize transfection efficiency, aim to generate results in the shortest possible time without the need for optimization or prefer to use a reagent proven many times over in practice, our product range supplies the ideal reagent for their individual needs and requirements.
Learn more about the transfection mechanism
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