Fig. 1.
Introducing the lux luciferin:luciferase operon components at 20–30:1 M ratios produces robust autobioluminescence in iPSCs. a Single operon, 2A-segmented, polycistronic lux operon driven by the chicken beta actin (CBA) promoter and flanked by sequence elements facilitating transposon-mediated genomic integration (TE). b Split lux cassette orientation enabling ratio-based component expression. F2A, foot and mouth disease viral 2A element; E2A, equine rhinitis A viral 2A element; Ta2A, synthetic Thosea asigna viral 2A element; P2A, Porcine teschovirus 1 viral 2A element; T2A, Thosea asigna viral 2A element. c Light production following transient transfection of Stem-luxCDEF:Stem-luxAB from b at a 1:1 M ratio compared to otherwise identical cells transfected with the same amount of Stem-luxAB but increasing molar ratios of Stem-luxCDEF. The average radiance was normalized to the MTT signal. Values are representative of N = 3 replicates. Error bars represent the standard error of the means. p/s/cm2/sr; photons/s/cm2/steradian. Data is available at https://osf.io/h5qzj/ [15]