CEACAM5 CRISPR Activation Plasmid (h): sc-401219-ACT

CEACAM5 (carcinoembryonic antigen-related cell adhesion molecule 5) is a glycosylphosphatidylinositol-anchored immunoglobulin superfamily protein expressed on the apical surface of epithelial cells, where it contributes to cell–cell adhesion and maintenance of tissue architecture. Through homophilic and heterophilic interactions, CEACAM5 can influence epithelial differentiation, polarity, and contact-dependent signaling that intersects with adhesion-dependent pathways and remodeling of the cell surface glycocalyx. Dysregulated CEACAM5 expression is frequently associated with epithelial malignancies and is widely used as a molecular marker in studies of tumor cell biology, including invasion, metastasis, and immune interactions within the tumor microenvironment. Its surface localization and variable expression across contexts make it a useful node for interrogating epithelial state changes and oncogenic signaling programs.

CEACAM5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CEACAM5 expression without altering the underlying DNA sequence.

CEACAM5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CEACAM5 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 CEACAM5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CEACAM5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CEACAM5 locus and enabling the study of CEACAM5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CEACAM5 pathway restoration in tumor cells with silenced or reduced CEACAM5 expression.

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