BCAM CRISPR Activation Plasmid (h): sc-405080-ACT

BCAM (basal cell adhesion molecule; Lutheran blood group glycoprotein) is a transmembrane immunoglobulin superfamily receptor that binds laminin α5-containing isoforms within basement membranes. By mediating cell–matrix adhesion, BCAM contributes to epithelial and endothelial interactions, cytoskeletal organization, and regulation of cell motility and tissue integrity. BCAM-associated adhesion signaling interfaces with extracellular matrix remodeling processes and can influence vascular and inflammatory microenvironments. Altered BCAM expression has been linked to dysregulated adhesion phenotypes observed in hematologic and solid tumor contexts, supporting its use as a marker and mechanistic node for studying disease-relevant cell trafficking and microenvironmental interactions.

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

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

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