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Methods Mol Biol ; 1742: 227-235, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29330804

RESUMO

Hypoxia has long been recognized as a driving force of tumor progression and therapeutic resistance, and the transcription factor HIF-1α is believed to play a crucial role in these processes. Here we describe an efficient RCAS/Nes-TVA system that allows for in vivo manipulation of HIF-1α expression in the mouse neural progenitor cells. Simple production of the recombinant avian virus RCAS enables quick delivery of gene of interest through injection into the neural progenitors of transgenic mice expressing the viral cognate receptor TVA under the nestin promoter. By crossing with various commercially available genetically engineered mouse strains, a repertoire of mouse models can be created to study gene-specific effects on glioma genesis. This chapter provides details of plasmid construction, viral production, and intracranial delivery of transgenes, a methodology that can be easily adapted to a specific purpose.


Assuntos
Carcinogênese/genética , Glioma/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Células-Tronco Neurais/citologia , Vírus do Sarcoma de Rous/genética , Animais , Proteínas Aviárias/genética , Hipóxia Celular , Linhagem Celular , Galinhas , Vetores Genéticos , Camundongos , Camundongos Transgênicos , Transplante de Neoplasias , Nestina/genética , Células-Tronco Neurais/patologia , Receptores Virais/genética
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