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
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author Pearson, Joel D.
Bremner, Rod
author_facet Pearson, Joel D.
Bremner, Rod
author_sort Pearson, Joel D.
collection PubMed
description 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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spelling pubmed-84935702021-10-08 Lentiviral-mediated ectopic expression of YAP and TAZ in YAP(off) cancer cell lines Pearson, Joel D. Bremner, Rod STAR Protoc Protocol 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). Elsevier 2021-10-01 /pmc/articles/PMC8493570/ /pubmed/34632420 http://dx.doi.org/10.1016/j.xpro.2021.100870 Text en Crown Copyright © 2021. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Pearson, Joel D.
Bremner, Rod
Lentiviral-mediated ectopic expression of YAP and TAZ in YAP(off) cancer cell lines
title Lentiviral-mediated ectopic expression of YAP and TAZ in YAP(off) cancer cell lines
title_full Lentiviral-mediated ectopic expression of YAP and TAZ in YAP(off) cancer cell lines
title_fullStr Lentiviral-mediated ectopic expression of YAP and TAZ in YAP(off) cancer cell lines
title_full_unstemmed Lentiviral-mediated ectopic expression of YAP and TAZ in YAP(off) cancer cell lines
title_short Lentiviral-mediated ectopic expression of YAP and TAZ in YAP(off) cancer cell lines
title_sort lentiviral-mediated ectopic expression of yap and taz in yap(off) cancer cell lines
topic Protocol
url 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
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