Mucin 2/MUC2 CRISPR Activation Plasmid (h): sc-400221-ACT

MUC2 encodes mucin 2, a major gel-forming secreted mucin that constitutes the structural backbone of the intestinal mucus layer and supports epithelial barrier integrity. Through extensive O-glycosylation and polymerization, MUC2 regulates mucosal hydration, microbial spatial organization, and protection from luminal stress, linking it to innate immune signaling and epithelial differentiation programs. Altered MUC2 expression or mucin architecture is associated with mucus barrier dysfunction, dysbiosis, and chronic inflammatory processes in the gastrointestinal tract. MUC2 is also widely used as a marker of goblet cell lineage and secretory epithelial states in studies of intestinal homeostasis and disease-related remodeling.

Mucin 2/MUC2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MUC2 expression without altering the underlying DNA sequence.

Mucin 2/MUC2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MUC2 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 MUC2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Mucin 2/MUC2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MUC2 locus and enabling the study of Mucin 2/MUC2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Mucin 2/MUC2 pathway restoration in tumor cells with silenced or reduced MUC2 expression.

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