Glucosidase I CRISPR Activation Plasmid (h): sc-405014-ACT

MOGS encodes glucosidase I, an ER-resident α-glucosidase that catalyzes the first glucose trimming step of N-linked glycan processing on nascent glycoproteins. This activity is central to ER quality control, influencing calnexin/calreticulin-assisted folding, secretion competence, and ER-associated degradation (ERAD) of misfolded proteins. By shaping glycoprotein maturation, MOGS impacts proteostasis and cellular responses to ER stress and the unfolded protein response (UPR). Altered MOGS function has been linked to congenital disorders of glycosylation and can modulate host–pathogen interactions through effects on viral envelope glycoprotein processing, supporting its relevance in glycobiology and infection-related research contexts.

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

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

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