Gabanyi I, Lojudice F H, Kossugue P M, Rebelato E, Demasi M A, Sogayar M C
Universidade de São Paulo, Centro de Terapia Celular e Molecular, Departamento de Bioquímica, Instituto de Química, São Paulo, SP, Brasil.
Braz J Med Biol Res. 2013 Feb;46(2):121-7. doi: 10.1590/1414-431x20122148. Epub 2013 Feb 1.
The type I herpes simplex virus VP22 tegument protein is abundant and well known for its ability to translocate proteins from one cell to the other. In spite of some reports questioning its ability to translocate proteins by attributing the results observed to fixation artifacts or simple attachment to the cell membrane, VP22 has been used to deliver several proteins into different cell types, triggering the expected cell response. However, the question of the ability of VP22 to enter stem cells has not been addressed. We investigated whether VP22 could be used as a tool to be applied in stem cell research and differentiation due to its capacity to internalize other proteins without altering the cell genome. We generated a VP22.eGFP construct to evaluate whether VP22 could be internalized and carry another protein with it into two different types of stem cells, namely adult human dental pulp stem cells and mouse embryonic stem cells. We generated a VP22.eGFP fusion protein and demonstrated that, in fact, it enters stem cells. Therefore, this system may be used as a tool to deliver various proteins into stem cells, allowing stem cell research, differentiation and the generation of induced pluripotent stem cells in the absence of genome alterations.
I型单纯疱疹病毒VP22包膜蛋白含量丰富,以其将蛋白质从一个细胞转运到另一个细胞的能力而闻名。尽管有一些报道对其转运蛋白质的能力提出质疑,将观察到的结果归因于固定假象或简单地附着于细胞膜,但VP22已被用于将多种蛋白质递送至不同细胞类型,引发预期的细胞反应。然而,VP22进入干细胞的能力问题尚未得到解决。我们研究了由于VP22能够内化其他蛋白质而不改变细胞基因组,它是否可以作为一种工具应用于干细胞研究和分化。我们构建了一个VP22.eGFP构建体,以评估VP22是否能够被内化并携带另一种蛋白质进入两种不同类型的干细胞,即成人牙髓干细胞和小鼠胚胎干细胞。我们生成了一种VP22.eGFP融合蛋白,并证明它实际上能够进入干细胞。因此,该系统可作为一种工具,在不改变基因组的情况下将各种蛋白质递送至干细胞,从而用于干细胞研究、分化以及诱导多能干细胞的产生。