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Biochem Biophys Res Commun ; 478(4): 1732-8, 2016 09 30.
Artículo en Inglés | MEDLINE | ID: mdl-27614311

RESUMEN

Various serotypes of adeno-associated virus (AAV) vectors have been used for gene therapy and as research tools. Among these serotypes, the AAV type 2 vector has been used successfully in human gene therapies. However, the transduction efficiency of AAV2 depends on the cell type, and this poses a problem in the efficacy of gene therapy. To improve the transduction efficiency of AAV2, we designed a small peptide consisting of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor peptide and the HIV-Tat sequence Tat-Y1068. Pre- or co-treatment of CYNOM-K1 cells from cynomolgus monkey embryo skin with Tat-Y1068 increased the transduction efficiencies in a dose-dependent manner and caused p38 phosphorylation. The transduction efficiency of AAV2 into the rat fibroblast cell line RAT-1 highly expressing EGFR was less than the transduction efficiency of AAV2 into CYNOM-K1 cells. Tat-Y1068 increased the transduction efficiency in RAT-1 cells in the same manner as in CYNOM-K1 cells. In conclusion, cell-permeable peptides possessing the EGFR tyrosine kinase inhibitor function might serve as a useful ingredient of AAV2 vector solution for increasing the transduction efficiency of gene therapies.


Asunto(s)
Péptidos de Penetración Celular/farmacología , Dependovirus/genética , Fibroblastos/efectos de los fármacos , Piel/efectos de los fármacos , Transducción Genética/métodos , Secuencia de Aminoácidos , Animales , Western Blotting , Línea Celular , Péptidos de Penetración Celular/síntesis química , Relación Dosis-Respuesta a Droga , Embrión de Mamíferos/citología , Fibroblastos/citología , Fibroblastos/metabolismo , Vectores Genéticos/genética , Macaca fascicularis , Microscopía Fluorescente , Fosforilación/efectos de los fármacos , Ratas , Reproducibilidad de los Resultados , Piel/citología , Piel/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo
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