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

Joel D. Pearson; Rod Bremner

STAR Protocols (Dec 2021)

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

Joel D. Pearson,
Rod Bremner

Affiliations

Joel D. Pearson: Lunenfeld Tanenbaum Research Institute, Mount Sinai Hospital, Sinai Health System, Toronto, ON M5G 1X5, Canada; Department of Ophthalmology and Vision Science, University of Toronto, Toronto, ON M5T 3A9, Canada; Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, Canada; Corresponding author
Rod Bremner: Lunenfeld Tanenbaum Research Institute, Mount Sinai Hospital, Sinai Health System, Toronto, ON M5G 1X5, Canada; Department of Ophthalmology and Vision Science, University of Toronto, Toronto, ON M5T 3A9, Canada; Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, Canada; Institute of Medical Science, University of Toronto, Toronto, ON M5S 1A8, Canada; Corresponding author

Journal volume & issue: Vol. 2, no. 4
p. 100870

Abstract

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Summary: 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 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).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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