EMT Reporter Models for Cancer Research: A Window into Invasion and Metastasis

Thu, November 12, 2020 12:00 PM - 1:00 PM EDT

Abstract:

Epithelial to mesenchymal transition (EMT) involves decreased cell adhesion and increased cell motility, marked by the downregulation of epithelial markers and upregulation of mesenchymal markers. Following this transition in real time can be challenging for breast cancer researchers, as in vitro EMT reporter models in mammary cell lines are not readily available. To address this need, ATCC scientists used CRISPR/Cas9 technology to engineer an EMT reporter into the MCF 10A mammary cell line. The resulting advanced model harbors a knock-in in which E-cadherin is fused with EmGFP for real-time monitoring of EMT progression. The transgene was verified at genomic DNA, transcript, and protein levels. Upon TGF-β stimulation, MCF 10A E-cadherin EmGFP shows enhanced migration, reduced E-cadherin-GFP expression, and elevated vimentin and fibronectin expression, making this engineered cell line an ideal model for screening anticancer therapeutics that target EMT.

Key Takeways:

  • ATCC scientists used CRISPR/Cas9 to engineer an E-cadherin EmGFP (ECAD-EmGFP) reporter knock-in into a breast epithelial cell line.
  • The integrity of the ECAD-EmGFP knock-in was verified at the genomic, mRNA, and protein level.
  • This advanced cancer model can be used to track EMT status of cells in vitro by monitoring GFP expression.


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