Embigin CRISPR Activation Plasmid (h): sc-408321-ACT

Human EMB encodes embigin, a type I transmembrane immunoglobulin superfamily glycoprotein that functions as an accessory subunit for monocarboxylate transporters and modulates cell–cell and cell–matrix interactions. By supporting lactate and pyruvate flux through MCT complexes, embigin contributes to metabolic homeostasis, pH regulation, and microenvironmental adaptation in proliferating tissues. EMB-linked biology intersects with pathways governing adhesion, migration, and nutrient utilization, processes frequently rewired in oncogenesis and inflammatory states. Dysregulated embigin expression has been reported in multiple disease-associated contexts, making it a useful node for mechanistic studies of metabolic remodeling and cellular invasiveness.

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

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

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