Despite intracellular molecular dynamics being fundamental to understand pathological, biomechanical or biochemical events, several processes are still not clear because of the difficulty of monitoring and measuring these phenomena. To engineer an effective fluorescent tool useful to improve protein intracellular tracking studies, we fused a supernegative green fluorescent protein, (−30)GFP, to a myogenic transcription factor, MyoD. The (−30)GFP-MyoD was able to pass the plasma membrane when complexed with cationic lipids. Fluorescence confocal microscopy showed the protein delivery in just 3 hours with high levels of protein transduction efficiency. Confocal acquisitions also confirmed the maintenance of the MyoD nuclear localization. To examine how the supernegative GFP influenced MyoD activity, we did gene expression analyses, which showed an inhibitory effect of (−30)GFP on transcription factor function. This negative effect was possibly due to a charge-driven interference m...

中文翻译：

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超阴性GFP在促进MyoD细胞内跟踪方面的优势和局限性

尽管细胞内分子动力学对于理解病理，生物力学或生化事件是必不可少的，但由于难以监测和测量这些现象，因此尚不清楚几个过程。为了设计一种有效的荧光工具，用于改善蛋白质细胞内跟踪研究，我们将（-30）GFP超负绿色荧光蛋白与成肌转录因子MyoD融合。（-30）GFP-MyoD与阳离子脂质复合后能够通过质膜。荧光共聚焦显微镜显示蛋白质在短短3小时内具有高水平的蛋白质转导效率。共焦收购也证实了MyoD核定位的维持。为了检查超阴性GFP如何影响MyoD活性，我们进行了基因表达分析，显示出（-30）GFP对转录因子功能的抑制作用。这种负面影响可能是由于电荷驱动的干扰...