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A transgenic rat with ubiquitous expression of firefly luciferase gene

机译:萤火虫荧光素酶基因无处不在的转基因大鼠

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摘要

In vivo imaging strategies provide cellular and molecular events in real time that helps us to understand biological processes in living animals. The development of molecular tags such as green fluorescent proteins and luciferase from the firefly Photinus pyralis has lead to a revolution in the visualization of complex biochemical processes. We developed a novel inbred transgenic rat strain containing firefly luciferase based on the transgenic (Tg) technique in rats. This Tg rat expressed the luciferase gene ubiquitously under control of the ROSA26 promoter. Cellular immune responsiveness against the luciferase protein was evaluated using conventional skin grafting and resulted in the long-term acceptance of Tg rat skin on wild-type rats. Strikingly, organ transplant with heart and small bowel demonstrated organ viability and graft survival, suggesting that cells from luciferase-Tg are transplantable to track their fate. Taking advantage of the less immunogenic luciferase, we also tested the role of hepatocyte-infusion in a liver injury model, and bone marrow-derived cells in a skin defect model. Employed in conjunction with modern advances in optical imaging, this luciferase-Tg rat system provides an innovative animal tool and a new means of facilitating biomedical research such as in the case of regeneration medicine.
机译:体内成像策略可实时提供细胞和分子事件,有助于我们了解活体动物的生物学过程。来自萤火虫萤火虫的分子标记,例如绿色荧光蛋白和萤光素酶的发展,导致了复杂生化过程可视化方面的一场革命。我们基于大鼠的转基因(Tg)技术开发了一种新型萤火虫自交系大鼠品系,其中含有萤火虫荧光素酶。该Tg大鼠在ROSA26启动子的控制下普遍表达萤光素酶基因。使用常规的皮肤移植评估了针对萤光素酶蛋白的细胞免疫应答,并导致Tg大鼠皮肤长期被野生型大鼠接受。令人惊讶的是,有心脏和小肠的器官移植证明了器官的生存能力和移植物的存活率,表明萤光素酶-Tg的细胞可移植以追踪其命运。利用免疫原性较低的荧光素酶,我们还测试了肝损伤模型中注入肝细胞的作用以及皮肤缺陷模型中骨髓衍生的细胞的作用。结合现代光学成像技术的应用,这种荧光素酶-Tg大鼠系统提供了创新的动物工具和促进生物医学研究的新手段,例如再生医学。

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