AGTRAS 1.0

Transfection is the process of introducing exogenous biological material into eukaryotic cells, in particular in mammalian cells. Usually genetic material such as DNA and siRNA are inserted, but also proteins (such as antibodies) can be transfected. The process and methods to implement it are analogous to those for bacterial transformation, which however concerns bacteria and, sometimes, plant cells.

The transfection process can be done:

1. in vitro - on target cells in long-term cell cultures;
2. ex vivo - on cells isolated from an organism and transferred onto culture medium;
3. in vivo - directly on cells of an organism.

Transfection is useful in all genetics and molecular biology studies in order to obtain information on the activity and function of certain genes. This can generally be done in two ways: overexpressing the gene or silencing it. In the first case a higher than normal production of the gene product is induced by inserting further copies of the same gene; in the second case, the gene will be "blocked". There are two methods to silence a gene: through the formation of knock-out systems, which exchange the gene in the cell with an inactive one or by inserting siRNA (RNA interference).

Depending on the fate of the inserted vector, a distinction can be made between transient transfection and stable transfection. Transfection is transient when the vector remains in the cell as an extrachromosomal fragment, i.e. it does not integrate into the cellular genome; in this case the properties induced by the transfection remain for a short time, usually disappear before 72 hours. Transfection is stable, however, when the exogenous DNA is permanently integrated into the genome: the effect therefore remains for the entire life of the cell and will also be transmitted to the cells that derive from it. Transient transfection is certainly faster, easier and cheaper but obviously allows studies limited in time. Stable transfection is considered necessary whenever the gene effect is medium-to-long-term: for example, when studying genes that are thought to be related to the processes of cell differentiation.

One of the simplest and less expensive transfection method is the one using calcium phosphate, developed for the first time by S. Bacchetti and F.L. Graham in 1977. Another very effective biological method exploits liposomes, small lipid vesicles that incorporate DNA and are induced to enter with it in the cell by simulating the processes of cellular endocytosis. Another method involves the use of dendrimers, highly branched molecules that bind to DNA and transport it to the cell.

NEXT Genomics aims to develop and optimize a transfection protocol using lipofectamine, a widely used transfection agent. The goal is to produce a technology that can be easily applied by the customer to offer "custom" transfection services.