FIGURE 3 from Overcoming Xenoantigen Immunity to Enable Cellular Tracking and Gene Regulation with Immune-competent “NoGlow” Mice

NoGlow mice express eGFP and Luciferase without background fluorescence and bioluminescence. A, Diagram of the NoGlow construct. LoxP-flanked stop site prevents expression of the NoGlow construct in the absence of Cre recombinase. However, in the presence of Cre the stop site is recombined to yield a single transcript encoding rtTA3 and mutant GFP/Luc driven by the CAG promoter. B, Chimeric founder NoGlow animals were crossed to CMV (full-body) Cre animals to test activity and expression of the construct. Representative bioluminescence imaging of female C57Bl/6, CAG Luc-GFP, and F1 CMV Cre NoGlow littermates with or without the NoGlow construct reveal no background bioluminescence in CMV Cre NoGlow+ animals. C, Bioluminescent imaging of representative lungs/livers shows no background bioluminescence in NoGlow+ mice. D, Chromogenic Luciferase staining in livers from NoGlow+ and control mice confirms luciferase protein expression despite the lack of bioluminescence in B and C. E, Representative fluorescent imaging of male C57Bl/6, CAG Luc-GFP, and F1 CMV Cre NoGlow animals shows no background fluorescence in CMV Cre NoGlow+ mice, while immunofluorescent staining (F) for GFP in liver sections from CMV Cre NoGlow+ animals confirms that GFP protein can be detected upon antibody staining.