Reporter Gene

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Caty784
Caty784's picture
Reporter Gene

Hi.
I know how the reporter gene works in a transfected cell. My question for anyone who would know is, WHY does it work?
From the time you make the gene construct (with a gene of interest for a protein of interest, say, ErbB2) and until the cell culture starts to expresses that on the plasma membrane of the individual cell, there are a million things that can go wrong, but usually don't, at least in a large percentage of the cells. Why is that? Why is the cell, first of all, allowing that new stretch of DNA, which is exogenous, to be transcribed into mRNA and then translated into the respective protein (say beta-Gal)? Any papers in the literature about this that anyone might know about?
Thank you very much!!!

Omai
Omai's picture
This paper has some

This paper has some reasonably good answers to your question.
Enjoy.

http://www.aapsj.org/view.asp?art=aapsj0901009

Omai

Jason King
Jason King's picture
wrote:

wrote:

Reporter constructs work because we have been able to work out which elements present upstream of the translation start site are required for expression of the mRNA. Almost all of this knowledge was gained from analysing how viruses get their genes expressed in the host cell. They have had to evolve promoters that are short and strong enough to get enough of their genes expressed to allow new progeny virions to form before the host organism "notices" and does something about it.

As most reporter constructs are only required transiently, the fact that the cell does recognize them as foreign isn't too great a problem. However, scientists doing non-viral gene therapy spend a lot of time modifying their nucleic acid based vectors so that they persist for as long as possible.