GeneCopoeia offers genome-wide human, mouse and rat microRNA (miRNA) 3' UTR target clones in mammalian expression vectors with luciferase reporter gene. Both dual luciferase and single luciferase vectors are offered for performing assays on cell lysate or without destroying target cells respectively.
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 miRNA on target gene(s).
3' UTR sequences were obtained from public domain gene sequence databases and inserted downstream of the firefly or Gaussia luciferase reporter gene in their respective vector system, driven by SV40 promoter for expression in mammalian cells. A chimeric mRNA is transcribed consisting of the firefly or Gaussia luciferase and a 3' UTR target sequence. Thus, the study of mRNA-miRNA target interaction can be performed with a functional assay for either firefly or Gaussia luciferase.
miTarget 3' UTR clones are available as ready-to-transfect or with custom service options.
Choose the right luciferase vector depending on your assay requirements
Genome wide coverage of human (23,000), mouse (19,000) and rat (8,000)
Simple systematic design
The 3' UTR target sequence is fused downstream to either a firefly or Gaussia luciferase. Expression of the luciferase is suppressed when a miRNA complex binds specifically to the 3' UTR target
Convenient dual reporter vector
Single vector with dual reporters including one for regulatory detection and another for internal control and signal normalization
Eliminate the need for transfection of two separate reporter constructs
Easy quantification with optimized assay kits
Luciferase activities can be easily quantified using either cell lysate (firefly luciferase, non-secreted) or supernatant without destroying target cells (Gaussia luciferase, secreted
Figure 2. The inhibitory effect of miRNA on a target sequence(3' UTR) expression clone can be measured with GeneCopoeia Luc-Pair™ miR Luciferase Assay kit. HEK 293 cells were plated on a 6-well plate. On the second day, the cells were transfected with 1.0 µg of Lin28 3' UTR miRNA target sequence expression clone in pEZX-MT01 vector with fLuc (Product ID: HmiT019205) and 1.4 µg of miR-125a precursor miRNA expression clone in pEZX-MR03 vector with eGFP (Product ID: HmiR0309) and miRNA control clone as indicated in the figure. The cells were transferred to a 96-well plate 18 hours after transfection and cultured for another 24 hours. Both firefly luciferase and Renilla luciferase activities were measured and data was recorded on Victor II machine. Firefly luciferase activity was then normalized with Renilla luciferase activities in the same well.
