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What is stable transfection?
Unlike transient transfection, stable transfection can integrate the transfected DNA
into the cell genome and pass it to the progeny of the cell, thereby achieving the
long-term introduction of foreign DNA into the cell. Stable transfection can be used to
study the long-term effects of gene expression or to create cell lines with new
characteristics for further experiments, such as screening potential drug candidates in
How does stable transfection work?
The purpose of stable, long-term transfection is to isolate and propagate a single
clone containing transfected DNA that has been integrated into the genome of the
cell. One of the most reliable methods for selecting cells that stably express
transfected DNA is selective screening. Commonly used selection markers include
genes that are resistant to various selection drugs. Long-term continuous antibiotic
treatment of cells will only lead to the expansion of stably transfected cells, and
unstable cells will die due to a lack of antibiotic resistance.
An alternative strategy is to use vectors carrying essential genes that are defective in
each cell line. For example, CHO cells that lack dihydrofolate reductase (DHFR) gene
expression cannot survive without the addition of nucleosides. However, these cells,
after being stably transfected with DNA expressing the DHFR gene, will synthesize the
required nucleosides and survive. Another advantage of using DHFR as a marker is
that when cells are exposed to increasing doses of methotrexate, gene amplification
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of DHFR and related transfected DNA occurs, resulting