Stable Cell Line Transient Transfection

Stable expression of target gene in cell lines overcomes the low transfection efficiency of transient expression and produces more target proteins.

Stable cell line transient transfection.

Typical cell density ranges are as follows. Therefore to make stably transfected cell lines foreign dna has to be integrated into the genome of the cell line. However stable inheritance can be observed in nongenomic dna as well. The plasmid dna or other type of nucleic acid typically has a reporter gene that allows a scientist to monitor the expression of the reporter gene usually within 1 2 days post transfection.

In a subpopulation of transfected cells whether the desired effect is a stable or transient transfection the transfected genetic material will integrate into the genome. Key difference transient vs stable transfection transfection is a process which is involved in the gene transfer of eukaryotic cells using chemical or physical methods. Stably transfected cells pass the introduced dna to their progeny typically because the transfected dna has been incorporated into the genome but sometimes via stable inheritance of nongenomic dna. When developing a stable transfection researchers use selectable markers to distinguish transient from stable transfections.

Stable transfection is the long term introduction of foreign dna into cells. Unlike the short term protein expression observed using transient transfection approaches generating cell lines using lentiviral vectors enables long term protein expression studies. This protocol can be used to generate stable cell lines expressing a gene of interest from an integrated lentiviral vector. In transient transfection the transfected material enters the cell but does not get integrated into the cellular genome.

Stably transfected cells pass the foreign dna through progeny. There are two types of stable cell lines. Transient transfection of mammalian cells has been employed for recombinant protein expression since the invention of transfection reagents the ability to express milligram to gram amounts of recombinant protein has relied mainly on the creation of stable cell lines. Transfection can be classified into main two types named transient transfection and stable transfection during transient transfection the gene of interest fails to integrate with the host genome and is expressed temporally.

0 8 2 4 10 5 cells ml of complete media. In order to create stable cell lines investigators will take advantage of this natural occurrence and introduce the gene of interest along with a selectable marker. One is achieved by eukaryotic vectors that harbor elements for episomal maintenance in the nucleus of a transfected cell.