DPAGT1 Double Nickase Plasmid (h): sc-406355-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DPAGT1 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
- DPAGT1 Double Nickase Plasmid (h) and DPAGT1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting DPAGT1. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DPAGT1 Double Nickase Plasmid (h) | sc-406355-NIC | 20 µg | $410.00 | |||
DPAGT1 Double Nickase Plasmid (h2) | sc-406355-NIC-2 | 20 µg | $410.00 |
DPAGT1 encodes dolichyl-phosphate N-acetylglucosaminephosphotransferase 1, an endoplasmic reticulum membrane enzyme that catalyzes the first committed step of N-linked glycosylation by transferring N-acetylglucosamine-1-phosphate to dolichol phosphate. This activity initiates lipid-linked oligosaccharide biosynthesis and supports co-translational protein folding and quality control within the ER, integrating with ER homeostasis and proteostasis pathways. Disruption of DPAGT1-dependent glycosylation perturbs maturation of many secretory and membrane proteins, influencing receptor trafficking and cell-surface glycoprotein composition. Pathogenic variants in DPAGT1 are linked to congenital disorders of glycosylation and DPAGT1-associated congenital myasthenic syndromes, making it a useful target for mechanistic studies of glycoprotein biogenesis and ER function.
DPAGT1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the DPAGT1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within DPAGT1. 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 DPAGT1 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 DPAGT1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.