3'UTR constructs - any available commercially (Oct/26/2009 )
We are a little tight on time right now, trying to roll a study. I have a few miRs of interest all predicted to target the same gene. Cloning the 3'UTR and then performing the studies might take a while. A quick google search gives 2 companies- GeneCopoeia and SwitchGear, that presumably sell pre-made 3'UTR constructs tagged to Luciferase reporter for my gene of interest.
I haven't noticed any publications citing their constructs. Just wondering if anyone had any experience with either of these 2 companies. Or if you knew any other that worked well for you.
Also, a related question, most Luc-reported studies use a microRNA expression plasmid co-transfected with their Luc-reporter plasmid. I was wondering if there is any reason that co-transfection with miR-mimics (eg: Pre-miRs from Ambion) are not popular for repression of Luciferase signal?
Any help is appreciated. Thanks!
In general, the 3'UTR is in one exon and it should not be a big problem to use PCR to pull out from genomic DNA.
Regarding your co-transfection question, most people might want to generate stable lines to constitutively express miRNA, so they use vector for co-transfection. Also, it's easy to monitor the transfection efficiency with markers, e.g. GFP and RFP, on the miRNA expression vector.