I don’t know wich sequence of miRNA must be cloned

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Erik Diaz

You need the actual gene without introns, you could include also the 5'utr region.

At the end you want to have the CDS to express it under a 'x' promoter.

Erika Girardi

I would recommend you to clone the pre-miR sequence +/- 100 nucleotides (100nt upstream and downstream of Drosha cleavage site at the base of the stem of the pre-miR). By doing so, you are on the safe side to have a good pri-miRNA substrate for Drosha cleavage. After this first processing, your pre-miRNA will be cleaved by Dicer and then your microRNA of interest will be loaded in Ago. You can check Narry Kim's papers for more informations.

Noor Idan Jarad

hi i agree with dr.Erika Girardi

Mohsen Saravani

Thank you all for providing useful information.