4 replies to this topic

[Priya914]

Hi all,
I am writing a proposal for my preliminary exam and I have a question. I am very new to this field of micro-RNA.
I wanted to know if I tested that my micro-RNA targets a gene in HeLa cells by luciferase assay, will the micro-RNA surely target this gene in my cell line of interest.
In case of HeLa cells, I will be transfecting the synthetic micro-RNA too. However, in my cell line of interest HBMVEC, I know this micro-RNA is expressed. So should I also do a luciferase report analysis in HBMVEC along with Hela cells?
I appreciate your help and time.
Thanks a lot.  

[Functional Screens]

The endogenous miRNA in HBMVEC cells might not be enough and sufficiently suppress the overexpressed luciferase reporter introduced by transfection.  That's why people co-transfect miRNA-expression vector and luciferase reporter in model cell lines such as 293T.

[Priya914]

Functional Screens, on Oct 27 2009, 12:05 PM, said:

The endogenous miRNA in HBMVEC cells might not be enough and sufficiently suppress the overexpressed luciferase reporter introduced by transfection.  That's why people co-transfect miRNA-expression vector and luciferase reporter in model cell lines such as 293T.

Thanks so much .  That answers my question partially. But do you think a micro-RNA targets different genes in different cell lines ?

[Functional Screens]

Due to mRNA expression profile with tissue specificity, I will say same miRNA might target different genes and give different phenotypes.  So, one miRNA can be a tumor suppressor in one cell line but acts as oncogene in another one.

[Priya914]

Thanks so much. So this implies I will have to check whether this miRNA targets the gene in both HeLa and HBMVEC.