Applications
As an experimentally transformed cell line, HEK 293 cells are not a particularly good model for normal cells, cancer cells, or any other kind of cell that is a fundamental object of research. However, they are extremely easy to work with, being straightforward to culture and to transfect, and so can be used in experiments in which the behavior of the cell itself is not of interest. Typically, these experiments involve transfecting in a gene (or combination of genes) of interest, and then analyzing the expressed protein; essentially, the cell is used simply as a test tube with a membrane. The widespread use of this cell line is due to its extreme transfectability by the various techniques, including calcium phosphate method, achieving efficiencies approaching 100%.
An important variant of this cell line is the 293T cell line that contains, in addition, the SV40 Large T-antigen, that allows for episomal replication of transfected plasmids containing the SV40 origin of replication. This allows for amplification of transfected plasmids and extended temporal expression of the desired gene products. Note that any similarly modified cell line can be used for this sort of work; HeLa, COS and Chinese Hamster Ovary cell are common alternatives.
Examples of such experiments include:
- A study of the effects of a drug on sodium channels
- Testing of an inducible RNA interference system
- Testing of an isoform-selective protein kinase C agonist
- Investigation of the interaction between two proteins
- Analysis of a nuclear export signal in a protein
A more specific use of HEK cells is in the propagation of adenoviral vectors. Viruses offer an extremely efficient means of delivering genes into cells, since this is what they have evolved to do, and are thus of great use as experimental tools. However, as pathogens, they also present a degree of danger to the experimenter. This danger can be avoided by the use of viruses which lack key genes, and which are thus unable to replicate after entering a cell. In order to propagate such viral vectors, a cell line that expresses the missing genes is required. Since HEK cells express a number of adenoviral genes, they can be used to propagate adenoviral vectors in which these genes (typically, E1 and E3) are deleted, such as AdEasy.
293, and especially 293T, cells are commonly used for the production of various retroviral vectors. Various retroviral packaging cell lines are based on these cells.
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