Figure 5
From: Development of a new caged intein for multi-input conditional translation of synthetic mRNA
Translational repression by the reconstituted EGFP- and eDHFR-responsive C46-split CaVT via orthogonal caged split inteins that enable OR gate. (a) Schematic and truth table of mRNAs used in target protein-responsive translational repression via orthogonal caged split intein. (b) Cell transfection using orthogonal caged split inteins that enable OR gate. HeLa cells were co-transfected with 2xScMS2(C)-Luc2 (10 ng/well), Nluc (1 ng/well), MS2CP(1–45)-eNpuNcage-Nb113 (17.5 ng/well), CA1698-NpuCcage-MS2CP(46–116)-VPg(FCV) (17.5 ng/well), MS2CP(1–45)-RmaN(-4)cage-Lag16 (17.5 ng/well), GFPenhNb-RmaC-MS2CP(46–116)-VPg(FCV) (17.5 ng/well), and EGFP or eDHFR (10 ng/well) mRNAs, also human codon-optimized Barstar mRNA (10 ng/well) as negative target mRNA. The bar graph shows the Luc2/Nluc ratio (mean ± SD, n = 4). *P < 0.05, **P < 0.01, ***P < 0.001 compared with the negative control by Tukey’s multiple comparison test.
