3'UTR luciferase assay with 5 miRNA target sites - how to design 5 miRNA target sites in the 3'UTR (Mar/17/2011 )

I would like to design a 3'UTR with 5 miRNA target sites for a luciferase assay. I need to know how close the target sites can be. Does anyone have experience with this?

Thanks!

PBody

I guess there is no definite rule regarding how close the targets should be. You can put them next to each other or add a few based in between. Since you have 5 targets, and you are going to synthesize oligos to create the insert, there is no much room for you to add many extra bases between targets.

peebee on Thu Mar 17 16:00:59 2011 said:

My favorite miRNA supplier did a similar custom job for me, but the journal wanted more definitive 1:1 proof first. I'd start with these: "http://www.abmgood.com/miRNA(microRNA)/3'UTR-miRNA-Reporters.html"