Fig. 3: Characterization of de novo luciferase activity in vitro and in human cells. | Nature

Fig. 3: Characterization of de novo luciferase activity in vitro and in human cells.

From: De novo design of luciferases using deep learning

Fig. 3: Characterization of de novo luciferase activity in vitro and in human cells.

a, Substrate concentration dependence of LuxSit, LuxSit-f and LuxSit-i activity. Numbers indicate the signal-to-background (S/N) ratio at Vmax (photon s−1 molecule−1). Data are mean ± s.d. (n = 3). b, Luminescence images acquired by a BioRad Imager (top) or an Apple iPhone 8 camera (bottom). Tubes from left to right: DTZ only; DTZ plus 100 nM purified LuxSit; and DTZ plus 100 nM purified LuxSit-i, showing the high efficiency of photon production. c, Fluorescence and luminescence microscopic images of live HEK293T cells transiently expressing LuxSit-i-mTagBFP2; LuxSit-i activity can be detected at single-cell resolution. Left, fluorescence channel representing the mTagBFP2 signal. Right, total luminescence photons were collected during a course of a 10-s exposure without excitation light, immediately after adding 25 µM DTZ. Insets, negative control, untransfected cells with DTZ. Scale bars, 20 μm; 40× magnification.

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