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Braz. j. med. biol. res ; 46(2): 121-127, 01/fev. 2013. graf
Article Dans Anglais | LILACS | ID: lil-668771

Résumé

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.


Sujets)
Animaux , Humains , Souris , Protéines de transport/pharmacocinétique , Membrane cellulaire/métabolisme , Cellules souches embryonnaires/métabolisme , Protéines à fluorescence verte/pharmacocinétique , Protéines virales structurales/pharmacocinétique , Technique de Western , Pulpe dentaire/cytologie , Cytométrie en flux , Protéines à fluorescence verte/génétique , Microscopie confocale , RT-PCR , Protéines virales structurales/génétique
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