Mucin 12/MUC12 CRISPR Activation Plasmid (h): sc-400505-ACT

MUC12 encodes mucin 12, a membrane-tethered, heavily O-glycosylated mucin that contributes to the epithelial glycocalyx and supports barrier function at mucosal surfaces. By shaping the pericellular microenvironment, MUC12 can influence cell–cell and cell–matrix interactions, apical membrane organization, and responses to inflammatory stimuli in gastrointestinal and other epithelia. Altered mucin expression and glycosylation are frequently associated with epithelial remodeling, dysregulated differentiation, and changes in adhesion and signaling programs. Accordingly, MUC12 is studied as a context-dependent marker and modulator in mucosal inflammation and epithelial cancers, including investigations of how mucin biology affects tumor–microenvironment interactions.

Mucin 12/MUC12 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MUC12 expression without altering the underlying DNA sequence.

Mucin 12/MUC12 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MUC12 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 MUC12 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Mucin 12/MUC12 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MUC12 locus and enabling the study of Mucin 12/MUC12-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Mucin 12/MUC12 pathway restoration in tumor cells with silenced or reduced MUC12 expression.

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