Real-time imaging of integrin ?4 dynamics using a reporter cell line generated by Crispr/Cas9 genome editing.

ABSTRACT: 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 ?4 subunit of the ?6?4 integrin. A tdTomato tag was inserted with a linker at the C-terminus of integrin ?4 in mouse mammary epithelial cells. Cells harboring this tagged allele were similar to wild-type cells with respect to integrin ?4 surface expression, association with the ?6 subunit, adhesion to laminin and consequent signaling. These integrin ?4 reporter cells were transformed with YAP (also known as YAP1), which enabled us to obtain novel insight into integrin ?4 dynamics in response to a migratory stimulus (scratch wound) by live-cell video microscopy. An increase in integrin ?4 expression in cells proximal to the wound edge was evident, and a population of integrin ?4-expressing cells that exhibited unusually rapid migration was identified. These findings could shed insight into integrin ?4 dynamics during invasion and metastasis. Moreover, these integrin ?4 reporter cells should facilitate studies on the contribution of this integrin to mammary gland biology and cancer.This article has an associated First Person interview with the first author of the paper.

SUBMITTER: Elaimy AL 

PROVIDER: S-EPMC6703706 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature