Figure 3
From: Development of a new caged intein for multi-input conditional translation of synthetic mRNA
Conditional translational repression by the caged Rma DnaB intein-mediated reconstitution of EGFP-responsive C46-split CaVT. (a) Schematic depiction of mRNAs used in EGFP-responsive translational repression. (b-c) HeLa cells were co-transfected with 2xScMS2(C)-Luc2 (10 ng/well), Nluc (1 ng/well), and two types of mRNAs to express split CaVT per group (40 ng/well each), and EGFP or eDHFR (10 ng/well) mRNAs. The positive control for translational repression was MS2CP(1–45)-RmaN(-4) with RmaC-MS2CP(46–116)-VPg(FCV) group. The negative control for translational repression was MS2CP(1–45)-RmaN(-4)cage or MS2CP(1–45)-RmaN(-4)Scage with RmaCcage-MS2CP(46–116)-VPg(FCV) group. EGFP-responsive translational repression of Luc2 mRNA by RmaN(-4)cage combined with the cage-free RmaC (b). EGFP-responsive translational repression of Luc2 mRNA by RmaN(-4)Scage combined with the cage-free RmaC (c). The bar graph shows the Luc2/Nluc ratio (mean ± SD, n = 4). **P < 0.01, ***P < 0.001 compared with eDHFR-transfected (EGFP-untransfected) cells in each group by the unpaired two-sided Student’s t- test.
