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Lentiviral-mediated ectopic expression of YAP and TAZ in YAP^off cancer cell lines.

ABSTRACT: 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).

SUBMITTER: Pearson JD

PROVIDER: S-EPMC8493570 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature

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Lentiviral-mediated ectopic expression of YAP and TAZ in YAPoff cancer cell lines.

Pearson Joel D JD Bremner Rod R

STAR protocols 20211001 4

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 (YAPoff) 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 ...[more]

PMID: 34632420

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