3'UTR miRNA Target Clones
10% OFF* miTarget™ 3'UTR miRNA Target Clones
Our partner GeneCopoeia offers genome-wide human, mouse and rat microRNA (miRNA) 3' UTR target clones in mammalian expression vectors. miRNA 3' UTR target clones can be used for miRNA target identification and functional validation of predicted targets, or to study the regulatory effect of miRNAs on target genes.
miTarget™ 3'UTR miRNA Target Clones in pEZX-MT05 - Gaussia Luciferase & Secreted Alkaline Phosphatase
3' UTR sequences were obtained from public domain gene sequence databases and inserted downstream of a secreted Gaussia luciferase (GLuc) reporter gene, which is expressed under control of the SV40 promoter in mammalian cells. A chimeric mRNA is transcribed consisting of the GLuc and a 3' UTR target sequence. Thus, the study of mRNA-miRNA target interaction can be easily performed with a live cell assay for GLuc using only 10 µl of the cell culture medium. Besides using GLuc as the miRNA 3' UTR target reporter, a secreted Alkaline Phosphatase (SEAP) reporter driven by a CMV promoter is also cloned into the same vector (pEZX-MT05) and serves as the internal control. The dual-reporter vector system enables transfection normalization for accurate across-sample comparison.
miTarget™ 3'UTR miRNA Target Clones in pEZX-MT06 – Firefly Luciferase & Renilla Luciferase
In addition to the GLuc/SEAP dual-reporter vector system, 3' UTR sequences are also vailable in a Firefly/Renilla Duo-Luciferase reporter vector (pEZX-MT06).
Special Price* Assay Kits
For miRNA target validation using GeneCopoeia's pEZX-MT05 and pEZX-MT06 target clones, the Secrete-Pair™ Dual Luminescence Assay Kit and Luc-Pair™ Duo-Luciferase Assay Kit, respectively, are recommended.
*Expires 30 June 2019
How 3’UTR miRNA Target Clones Work
The inhibitory effect of miRNA on a 3' UTR target sequence expression clone
LIN28 is the known target gene for miR-125a. HEK 293 cells were transfected with LIN28 3' UTR target sequence expression plasmid alone (left bar) or co-transfected with miR-125a precursor expression plasmid (right bar). Both the GLuc activity and an internal control SEAP activity were determined 24 hours post-transfection. The activity ratio of GLuc to SEAP was set to 1 for the single transfection sample with LIN28 3'-UTR target sequence expression clone (left bar), against which the activity of the co-transfection sample was compared to. The result shows that miR-125a suppressed the luciferase activity from the GLuc-LIN28-3'-UTR clone by more than 70% (right bar).
Vector Maps and Compatible Assay Kits
All 3'UTR miRNA Target Clones are available in the vectors listed below. Configure your gene specific construct in our clone search:
|Vector||Reporter gene||Tracking gene||Assay Type||Compatible Assay Kit|
|pEZX-MT05||Gaussia luciferase (GLuc)||Secreted alkaline phosphatase (SEAP)||Live cell assays||Secrete-Pair™ Dual Luminescence Assay Kit|
|pEZX-MT06||Firefly luciferase||Renilla luciferase||Cell lysis required||Luc-Pair™ Luciferase Assay Kit|