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The ability to monitor changes in the expression and localization of integrins is essential for understanding their contribution to development, tissue homeostasis and disease. Here, we pioneered the use of Crispr/Cas9 genome editing to tag an allele of the β4 subunit of the α6β4 integrin. A tdTomato tag was inserted with a linker at the COOH-terminus of β4 in mouse mammary epithelial cells. Cells harboring this tagged allele were similar to wild-type cells with respect to β4 surface expression, association with the α6 subunit, adhesion to laminin and consequent signaling. These β4 reporter cells were transformed with YAP, which enabled us to obtain novel insight into β4 dynamics in response to a migratory stimulus (scratch wound) by livecell video microscopy. An increase in β4 expression in cells proximal to the wound edge was evident and a population of β4 expressing cells that exhibited unusually rapid migration was identified. These findings could shed insight into β4 dynamics during invasion and metastasis. Moreover, these β4 reporter cells should facilitate studies on the contribution of this integrin to mammary gland biology and cancer. [ABSTRACT FROM AUTHOR]

Cancer stem cells (CSCs) contribute to multidrug resistance, tumor recurrence and metastasis, making them prime therapeutic targets. Their ability to differentiate and lose stem cell properties makes them challenging to study. Currently, there is no simple assay that can quickly capture and trace the dynamic phenotypic changes on the CSC surface. Here, we report rapid discrimination of breast CSCs from non-CSCs using a nanoparticle-fluorescent-protein based sensor. This nanosensor was employed to discriminate CSCs from non-CSCs, as well as CSCs that had differentiated in vitro in two breast cancer models. Importantly, the sensor platform could also discriminate CSCs from the bulk population of cells in patient-derived xenografts of human breast cancer. Taken together, the results obtained demonstrate the feasibility of using the nanosensor to phenotype CSCs and monitor their fate. Furthermore, this approach provides a novel area for therapeutic interventions against these challenging targets., (Copyright © 2018 Elsevier Inc. All rights reserved.)