Figure 3.

mtRNR1- and AES-Containing 3′ UTRs Promote High Protein Expression

(A) 3′ UTR motives were cloned downstream of a standard luciferase reporter gene (Luc). After electroporation of the respective mRNAs with equal amounts into hDCs, luciferase activity was measured in relative light units (RLU) to obtain protein decay kinetics. (B) Based on these data, total protein expressed over time (the area under the curve) was calculated and is depictured relative to the hBg 3′ UTR-containing mRNA. One-way ANOVA, Dunnett’s post-test; ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05 (three independently performed experiments). (C and D) mRNA coding for luciferase with the indicated 3′ UTR motives was electroporated into murine myoblasts (C2C12) (C) or human foreskin fibroblasts (HFFs) (D), and, based on protein decay kinetics, total protein expressed over time was calculated and compared to mRNA containing hBg 3′ UTR, as described above. One-way ANOVA, Dunnett’s post-test; ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05 (three independently performed experiments).