Fig. 1: Design and construction of the LuminON transgene expression system.
a Schematic representation of the LuminON system. In the presence of furimazine or blue light, luminGAVPO homodimerizes, interacts with UASG elements (5xUASG) and initiates transgene expression of the gene of interest (GOI). b Schematic representation of the reporter plasmid containing 5xUASG upstream of the TATA box and the transactivator plasmids encoding different configurations of Nluc-GAVPO fusion proteins. c Furimazine (FZ)- or blue light-induced Gluc expression by different configurations of Nluc-GAVPO transactivators driven by the CMV or SV40 promoter. d Quantitative control of Gluc expression by modulating furimazine concentration and blue light intensity. The transfected cells were cultured with different furimazine concentrations (0–25 μM) or light irradiances (0–1.63 W/m2). Gluc activity was determined 24 h after induction, and induction ratios were calculated. e Gluc expression kinetics upon induction by furimazine for different durations. The transfected cells were induced by furimazine for different durations within 12 h, and Gluc activity was measured 24 h after the first induction. Data were normalized to Gluc expression in the cells kept in dark conditions without induction by furimazine. f LuminON system-mediated Gluc expression in different mammalian cell types. Data were normalized to Gluc expression simultaneously induced by furimazine and external blue light. Data in (c, e and f) represent the mean ± s.d. from three technical replicates. All the statistical comparison was performed by two-tailed t test. n.s. means not significantly different, *P < 0.05, **P < 0.01, ***P < 0.001. Source data are provided in the Source Data file.
