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Transfection is the process of deliberately introducing naked or purified nucleic acids into eukaryotic cells. [1] [2] It may also refer to other methods and cell types, although other terms are often preferred: "transformation" is typically used to describe non-viral DNA transfer in bacteria and non-animal eukaryotic cells, including plant cells.
A typical optical transfection protocol is as follows: [11] 1) Build an optical tweezers system with a high NA objective 2) Culture cells to 50-60% confluency 3) Expose cells to at least 10 μg/mL of plasmid DNA 4) Dose the plasma membrane of each cell with 10-40 ms of focussed laser, at a power of <100 mW at focus 5) Observe transient transfection 24-96h later 6) Add selective medium if the ...
The cost of growing mammalian cell cultures is high, so research is underway to produce such complex proteins in insect cells or in higher plants, use of single embryonic cell and somatic embryos as a source for direct gene transfer via particle bombardment, transit gene expression and confocal microscopy observation is one of its applications ...
Human embryonic kidney 293 cells, also often referred to as HEK 293, HEK-293, 293 cells, are an immortalised cell line derived from HEK cells isolated from a female fetus in the 1970s. [ 1 ] [ 2 ] The HEK 293 cell line has been widely used in research for decades due to its reliable and fast growth and propensity for transfection .
In cell-based therapies transfection is used for applications such as ex-vivo gene therapy, [14] hematopoietic stem cell engineering, [15] production of induced pluripotent stem cells, [16] and ex-vivo preparation of cells for immunotherapy [17] Over the last 50 years nucleic acid transfection has been the most common subcategory of ...
Magnet-assisted transfection is a relatively new and time-saving method to introduce nucleic acids into a target cell with increased efficiency. In particular, adherent mammalian cell lines and primary cell cultures show very high transfection rates. Suspension cells and cells from other organisms can also be successfully transfected.
Nucleofection is a method to transfer substrates into mammalian cells so far considered difficult or even impossible to transfect. Examples for such substrates are nucleic acids, like the DNA of an isolated gene cloned into a plasmid, or small interfering RNA (siRNA) for knocking down expression of a specific endogenous gene.
Transient expression, more frequently referred to "transient gene expression", is the temporary expression of genes that are expressed for a short time after nucleic acid, most frequently plasmid DNA encoding an expression cassette, has been introduced into eukaryotic cells with a chemical delivery agent like calcium phosphate (CaPi) or polyethyleneimine (PEI). [1]