Real-time imaging of integrin β4 dynamics using a reporter cell line generated by Crispr/Cas9 genome editing

J Cell Sci. 2019 Jul 31;132(15):jcs231241. doi: 10.1242/jcs.231241.


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 C-terminus of integrin β4 in mouse mammary epithelial cells. Cells harboring this tagged allele were similar to wild-type cells with respect to integrin β4 surface expression, association with the α6 subunit, adhesion to laminin and consequent signaling. These integrin β4 reporter cells were transformed with YAP (also known as YAP1), which enabled us to obtain novel insight into integrin β4 dynamics in response to a migratory stimulus (scratch wound) by live-cell video microscopy. An increase in integrin β4 expression in cells proximal to the wound edge was evident, and a population of integrin β4-expressing cells that exhibited unusually rapid migration was identified. These findings could shed insight into integrin β4 dynamics during invasion and metastasis. Moreover, these integrin β4 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.

Keywords: Crispr/Cas9; Integrin β4; Migration; YAP.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adaptor Proteins, Signal Transducing / genetics
  • Adaptor Proteins, Signal Transducing / metabolism
  • CRISPR-Cas Systems*
  • Gene Editing*
  • Humans
  • Integrin alpha6 / genetics
  • Integrin alpha6 / metabolism
  • Integrin beta4 / genetics
  • Integrin beta4 / metabolism*
  • Microscopy, Video
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • YAP-Signaling Proteins


  • Adaptor Proteins, Signal Transducing
  • ITGA6 protein, human
  • ITGB4 protein, human
  • Integrin alpha6
  • Integrin beta4
  • Transcription Factors
  • YAP-Signaling Proteins
  • YAP1 protein, human