Glucosidase IIβ Double Nickase Plasmid (h): sc-404394-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Glucosidase IIβ Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- Glucosidase IIβ Double Nickase Plasmid (h) and Glucosidase IIβ Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PRKCSH. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Glucosidase IIβ Antibody (H-4): sc-374457
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Glucosidase IIβ Double Nickase Plasmid (h) | sc-404394-NIC | 20 µg | $410.00 | |||
Glucosidase IIβ Double Nickase Plasmid (h2) | sc-404394-NIC-2 | 20 µg | $410.00 |
PRKCSH encodes the β-subunit of glucosidase II, an endoplasmic reticulum (ER) luminal enzyme complex that trims glucose residues from N-linked glycans during glycoprotein maturation. This processing step supports the calnexin/calreticulin quality-control cycle and helps coordinate ER-associated degradation (ERAD) for misfolded proteins, linking PRKCSH function to proteostasis and unfolded protein response signaling. Disruption of glucosidase IIβ activity can perturb secretion and surface expression of glycoproteins, altering cell signaling and stress adaptation. Genetic variation in PRKCSH has been associated with autosomal dominant polycystic liver disease, making it relevant for studies of epithelial biology, organelle stress, and glycoprotein-dependent pathways.
Glucosidase IIβ Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PRKCSH locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PRKCSH. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt PRKCSH function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of PRKCSH-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.