Skip to main content
. 2017 Feb 27;13(2):e1006623. doi: 10.1371/journal.pgen.1006623

Fig 7. Regulation of target gene sensitivity to miRNA suppression by 5’UTR.

Fig 7

(A) An engineered psiCheck2 vector (psiCheck-2-pd) for investigating the effect of 5’UTR and 3’UTR on reporter gene expression. TSS, transcription start site. (B) Experimental scheme of reporter assays in primary B cells. FACS plots show electroporation efficiency using a GFP-expressing plasmid. (C,D) Dual luciferase reporter assay to determine the effect of 5’UTR and 3’UTR on the reporter gene protein (luciferase activity) (C) and mRNA (qRT-PCR) levels (D). Closed and open circles indicate reporters with wild-type (wt) and mutated (mut) CD69 3’UTR, respectively. A comparison of renilla luciferase activity normalized to firefly luciferase activity (hRluc/Fluc) between psiCheck-2-pd containing mut and wt CD69 3’UTR reveals the sensitivity of the renilla luciferase mRNA (hRluc) to miR-17~92-mediated suppression. Results of normalized hRlcu/Fluc (n = 10) are from three independent experiments. Each experiment contained 3–4 replicates.