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[kaveh]

Hello,

I am planning to validate the effect of a Drosophila microRNA on destabilization of a transcript. The 3UTR of the transcript is a predicted target of that microRNA.
I need to clone the 3UTR of the transcript in the downstream of luciferase and use the vector to transfect Drosophila S2 cells.

My question:

1) What is the most common/affordable luciferase vector/system for Drosophila S2 cells?
Promega has the "pmirGLO vector with dual luciferases. However, they cannot confirm if the vector works in insect cells (the luciferase has PGK promoter, which I am not sure if it works in Drosophila cells)

2) What would you use for over-expression of microRNA in S2 cells?
My best bet as of now is the pAC5.1 vector with actin promoter.

any input is highly appreciated.

K