Target gene cloning - (Mar/07/2016 )

How are you ladies and gentlemen,

I am quite a newbie in this field of miRNA and I have a question.  I am trying to clone a plant miRNA and its target gene and further overexpress them in a plant species but I am not sure if for the target gene I only have to focus on the ORF or include the 3' UTR and the 5'UTR.  If the target site of my miRNA is in the ORF of its target gene.

Thank you!

Just the ORF should be fine.

You need the 3'-UTR if you want the target gene to be regulated by the miRNA.

Typically the miRNA response element (MRE) is in the 3'-UTR.  When the mRNA cyclizes before translation, the RISC bound to the 3'-UTR MRE is brought close to the 5'-cap and the assembling initiation complex.  It is the interaction of the RISC with an element of the initiation complex that supresses translation.

Hey there,

Thanks a lot I greatly appreciate.

Skovic16 on Tue Mar 8 02:06:21 2016 said:

  I am trying to clone a plant miRNA and its target gene and further overexpress them in a plant species

I'm confused, you mean you want to clone miRNA and its target gene, these two both in a plant species?