In transient transfection our desire plasmid enter into cytoplasm not in the nucleus. Then how from that cytoplasmic plasmid, mRNA produce and finally translation into protein? This is because, which polymerase enzymes are required for that steps all are present in nucleus. So how does transient transfection actually work?
Hi,
Actually, the plasmid does enter the nucleus and is subsequently transcribed by nuclear RNA polymerases.
The plasmid you constructed does not have the ability to cross the cell membrane but remains tits ability to enter the nucleus. After your transfection (can be done by various method), the plasmid will enter the nucleus and directly transcribed without being integrated into the genome (in the case your plasmid has the ability of genome integration).
Hi
Sometimes you are going to transfect a prokaryotic cell which in this case there will be no barriers on the plasmid,s way to reach the enzyme.
But in the case of a eukaryotic cell, after entrance to the cytoplasm, the plasmid will use the intracytoplasmic transport machinery and the proteins located on nuclear pores to enter the nucleus.
plasmid always goes to the nucleus, to produce mRNA and then eventually protein. transient= the effect does not last long, its good for few days
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