GBE1 CRISPR Activation Plasmid (m): sc-428773-ACT

Mouse Gbe1 encodes glycogen branching enzyme 1 (GBE1), a key glycosyltransferase that introduces α-1,6 branch points during glycogen biosynthesis. By controlling glycogen architecture, GBE1 supports normal glucose storage, glycogen solubility, and efficient mobilization in metabolically active tissues, intersecting with core carbohydrate metabolism and energy homeostasis pathways. Perturbation of glycogen branching alters glycogen particle structure and can promote accumulation of poorly branched, amylopectin-like polysaccharides, making Gbe1 a relevant node for studying glycogen quality control and metabolic stress responses. Dysregulated GBE1 activity is associated with glycogen storage pathology and neuromuscular phenotypes in model systems, supporting its use in mechanistic studies of glycogen metabolism–linked disease biology.

GBE1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Gbe1 expression without altering the underlying DNA sequence.

GBE1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Gbe1 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 Gbe1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GBE1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Gbe1 locus and enabling the study of GBE1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GBE1 pathway restoration in tumor cells with silenced or reduced Gbe1 expression.

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