How to conclude microRNA induction leading to upregulation of 3'UTR-reporter luciferase assay ?

07 November 2017 1 6K Report

Hello everyone,

I am experiencing some weird output during my experiments which is as follows : -

I am using 3'UTR-reporter luciferase assay to assess the effect of microRNA of my interest but, instead of suppression I am getting an increase of around 40-50% in the luciferase intensity.

To tackle this I made sure that I am using similar amount of pMIR-report vector in each well by making a master mix of my pMIR vector along with OPTI and plus.

But still the results are almost consistent.

Can anyone please help me to resolve this issue ?

Could it be just an artifact or a believable readout ? Has anyone experienced such a thing before ?

Vasu Saini

Hello Kamil,

thank you for your thoughts,

I have been using an expression vector to express my miRNA of interest (not mimics).

Simultaneously I am trying to screen few other microRNAs for possible interactions which shows no effect (neither positive nor negative) which should serve as a control in a way.

thats what made me more confused about this set of data.

any comments would be more than welcome

thanks in advance

Best

Vasu

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