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Cell Transfection

Transfection of a GFP-expressing plasmid
Transfection of THP-1 cells
with a GFP-expressing plasmid

Cell transfection consists of introducing ‘foreign’ nucleic acids, such as plasmids or siRNAs, into a cell to either overexpress or to silence a gene of interest, respectively. Transfection allows for the functional evaluation of the expression of specific genes in a variety of cellular contexts. Depending on the type of nucleic acid, as well as the cell type, a number of different transfection protocols can be used. The three main methods of transfection are chemical (e.g. cationic lipids), physical (e.g. electroporation), and biological (e.g. viral transduction). The newly introduced nucleic acids can exist either transiently, whereby the gene is only expressed for a limited period of time and does not replicate, or stably, whereby the gene integrates into the genome of the recipient cell. After cell transfection, a key factor in the stable incorporation of a gene into the genome is the maintenance of selection pressure in cell culture using an appropriate selective antibiotic. 

Still hard-to-transfect cells, such as immune cells, often express potent cytosolic sensors that detect foreign nucleic acids. Therefore, a transfection enhancer or booster is required to disable these defenses, helping facilitate the expression of nucleic acids and greatly improve transfection success.

 

InvivoGen offers:

  • LyoVec™: A proprietary cationic lipid-based transfection reagent belonging to a family of phosphonolipids. It is simple to use in the transfection of numerous cell lines, such as the human embryonic kidney -293 (HEK-293) cell line.
     
  • NATE™: A nucleic acid transfection enhancer to boost both transient and stable transfection efficiencies. It has been designed for use in combination with transfection reagents and techniques recommended for hard-to-transfect cells, such as human monocytes and murine macrophages (i.e. THP-1 and RAW 264.7, respectively). 
     
  • Selective Antibiotics: A comprehensive range of highly pure and endotoxin-free selective antibiotics including, Blasticidin, Zeocin™, Puromycin, G418, Hygromycin B Gold, and Phleomycin. They can be used for selection in both mammalian and bacterial cells.

 

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