Glucosidase IIα Double Nickase Plasmid (h): sc-407683-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Glucosidase IIα 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 IIα Double Nickase Plasmid (h) and Glucosidase IIα Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GANAB. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Glucosidase IIα Antibody (D-1): sc-518226
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Glucosidase IIα Double Nickase Plasmid (h) | sc-407683-NIC | 20 µg | $410.00 | |||
Glucosidase IIα Double Nickase Plasmid (h2) | sc-407683-NIC-2 | 20 µg | $410.00 |
GANAB encodes the alpha subunit of glucosidase II, an ER-resident enzyme that sequentially trims glucose residues from N-linked glycans on nascent glycoproteins. This deglucosylation step is central to ER quality control, coupling glycan processing to calnexin/calreticulin-assisted folding and targeting persistently misfolded proteins to ER-associated degradation. Through these processes, GANAB influences secretory pathway proteostasis, unfolded protein response signaling, and maturation of many membrane and secreted proteins. Genetic and functional perturbation of GANAB has been linked to disorders of glycoprotein processing and has been studied in the context of polycystic kidney and liver disease mechanisms and broader ER stress–associated phenotypes.
Glucosidase IIα Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GANAB locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GANAB. 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 GANAB 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 GANAB-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.