Ep-CAM CRISPR Activation Plasmid (m): sc-421499-ACT

Mouse Epcam encodes Ep-CAM, a calcium-independent cell–cell adhesion glycoprotein enriched in epithelial tissues that helps organize adherens junctions and regulate epithelial architecture. Beyond adhesion, Ep-CAM participates in signaling networks that influence proliferation and differentiation, including crosstalk with Wnt/β-catenin–associated transcriptional programs and modulation of epithelial–mesenchymal dynamics. Altered EPCAM expression is linked to changes in barrier integrity and epithelial plasticity that are frequently studied in developmental biology, tissue regeneration, and oncology models. In mouse systems, Epcam is commonly used as a marker and functional regulator of epithelial lineage identity in contexts such as organoid biology and tumor progression.

Ep-CAM CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Epcam expression without altering the underlying DNA sequence.

Ep-CAM CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Epcam locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the Epcam transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ep-CAM expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Epcam locus and enabling the study of Ep-CAM-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ep-CAM pathway restoration in tumor cells with silenced or reduced Epcam expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.