Open AccessReal-time imaging of integrin β4 dynamics using a reporter cell line generated by Crispr/Cas9 genome editing
Author(s) -
Ameer L. Elaimy,
Mengdie Wang,
Ankur Sheel,
C. Mackenzie Brown,
Melanie R. Walker,
John J. Amante,
Wen Xue,
Alice Chan,
Christina E. Baer,
Hira Lal Goel,
Arthur M. Mercurio
Publication year - 2019
Publication title -
journal of cell science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.384
H-Index - 278
eISSN - 1477-9137
pISSN - 0021-9533
DOI - 10.1242/jcs.231241
Subject(s) - biology , integrin , microbiology and biotechnology , crispr , genome editing , cell migration , live cell imaging , population , cell adhesion , cell culture , wound healing , cell type , cell , gene , genetics , demography , sociology
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 live-cell 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.