Mucin 16/MUC16 CRISPR Activation Plasmid (h): sc-403050-ACT

MUC16 encodes mucin 16, a high–molecular weight, heavily O-glycosylated transmembrane mucin that contributes to the epithelial glycocalyx and mucosal barrier function. Through its extensive extracellular tandem repeats and cytoplasmic tail, MUC16 modulates cell–cell and cell–matrix interactions, impacts adhesion and migration programs, and can influence receptor signaling networks and immune recognition at the cell surface. Altered MUC16 expression and glycosylation are widely studied in the context of epithelial malignancies, where they correlate with changes in invasion, metastatic potential, and tumor microenvironment interactions. As such, MUC16 is frequently used as a functional node to interrogate surface mucin biology, glyco-epitopes, and membrane-proximal signaling in human cell models.

Mucin 16/MUC16 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MUC16 expression without altering the underlying DNA sequence.

Mucin 16/MUC16 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MUC16 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 MUC16 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Mucin 16/MUC16 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MUC16 locus and enabling the study of Mucin 16/MUC16-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Mucin 16/MUC16 pathway restoration in tumor cells with silenced or reduced MUC16 expression.

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