Fig. 6: Orthogonal aptazyme switch-mediated gene regulation in HEK293T cells.

a Schematic illustrating 3’-UTR aptazyme-mediated conditional regulation of mRNA stability. DHEF ligand binding induces an active conformation in the ribozyme, leading to its activation and subsequent mRNA destabilization. GOI and (A)n denote the gene of interest and the poly-A tail, respectively. b Fluorescence microscopy images of HEK293T cells transiently transfected with the aptazyme-regulated mCherry-BFP dual reporter. Cells were treated with the indicated DHEF concentrations and incubated at 37 °C for 48 h before imaging, scale bar: 50 μm. c Relative mCherry/BFP fluorescence intensity ratios at the indicated DHEF concentrations (0 to 200 µM) (n = 3 biological replicates); the statistical significance was determined by one-way analysis of variance (ANOVA), followed by Dunnett’s multiple comparisons test. d RT-qPCR analysis of relative mCherry mRNA levels in HEK293T cells under untreated and DHEF-treated conditions (n = 3 biological replicates); the statistical significance was determined by the two-tailed t-test. e Immunoblot analysis of mCherry protein levels in HEK293T cells with and without DHEF treatment. β-Actin served as a loading control (left panel). The graph (right panel) shows Western blot quantification normalized to β-Actin (n = 3 biological replicates); the statistical significance was determined by the two-tailed t-test. For all the relevant panels, data are shown as mean ± standard deviation (SD). Source Data are provided as a Source Data file.