Maltase-glucoamylase CRISPR Activation Plasmid (h): sc-404265-ACT

Human MGAM encodes maltase-glucoamylase, an intestinal brush-border α-glucosidase that hydrolyzes terminal α-1,4-linked glucose residues from maltose, maltotriose, and maltooligosaccharides to release free glucose. This enzyme acts in the final steps of dietary starch and glycogen digestion, functionally complementing sucrase-isomaltase and shaping postprandial carbohydrate availability at the mucosal surface. MGAM expression and activity intersect with enterocyte differentiation programs and nutrient-sensing networks that influence epithelial metabolism. Genetic or acquired reductions in MGAM function have been associated with carbohydrate malabsorption phenotypes and are relevant to studies of intestinal dysfunction and metabolic homeostasis.

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

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

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