Lentiviral-mediated ectopic expression of YAP and TAZ in YAP(off) cancer cell lines

Ectopic/overexpression systems are important for studying protein function, but care must be taken to avoid artifacts due to excessively high levels of overexpression. To study the function of YAP/TAZ in YAP/TAZ-deficient (YAP(off)) cancers, we developed a lentiviral system using weak, constitutive...

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Detalles Bibliográficos
Autores principales: Pearson, Joel D., Bremner, Rod
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8493570/
https://www.ncbi.nlm.nih.gov/pubmed/34632420
http://dx.doi.org/10.1016/j.xpro.2021.100870
Descripción
Sumario:Ectopic/overexpression systems are important for studying protein function, but care must be taken to avoid artifacts due to excessively high levels of overexpression. To study the function of YAP/TAZ in YAP/TAZ-deficient (YAP(off)) cancers, we developed a lentiviral system using weak, constitutive promoters to ectopically express YAP/TAZ to physiologically relevant levels. We detail this system along with protocols to assess YAP/TAZ expression by flow cytometry and quantitative western blotting. This system can also be easily adapted for the study of other proteins. For complete details on the use and execution of this protocol, please refer to Pearson et al. (2021).

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