Difference between revisions of "Team:Freiburg/Methods"

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<p>Polyethylenimine (PEI) is used as a polymeric transfection. PEI condenses DNA into positively charged particles, which bind to anionic cell surface residues and are brought into the cell via endocytosis. Using this method transient transfected HEK cells were generated.</p>
 
<p>Polyethylenimine (PEI) is used as a polymeric transfection. PEI condenses DNA into positively charged particles, which bind to anionic cell surface residues and are brought into the cell via endocytosis. Using this method transient transfected HEK cells were generated.</p>
 
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<h2>Electroporation</h2>
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<p>Electroporation is a method, which is used to introduce target genes into cells transiently in a physical way. To achieve this, an electrical field is generated around the cells to increase the permeability of the cell membrane, so that DNA can enter the cells. During our project we used electroporation to generate transient transfected Jurkat cells.</p>
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Revision as of 13:28, 12 October 2017


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Cell Culture

Transfection

Mammalian cell transfection is used to introduce foreign nucleic acids into cells. Depending on cell line and desired effect, there are different ways to transfect mammalian cells. These methods can be broadly classified into two groups: transient and stable transfections. Normally, a selection marker is needed to identify cells that are transfected.

Transient Transfection

Transient transfections generate short-term impact of alterations in gene and protein expression. In this case, the expression of the introduced gene is temporary because the foreign nucleic acids are not integrated into the host cell genome and therefore will not be passed on by cell division. Transient transfection of cells was achieved by two different methods:

PEI transfection

Polyethylenimine (PEI) is used as a polymeric transfection. PEI condenses DNA into positively charged particles, which bind to anionic cell surface residues and are brought into the cell via endocytosis. Using this method transient transfected HEK cells were generated.

Electroporation

Electroporation is a method, which is used to introduce target genes into cells transiently in a physical way. To achieve this, an electrical field is generated around the cells to increase the permeability of the cell membrane, so that DNA can enter the cells. During our project we used electroporation to generate transient transfected Jurkat cells.