AGA Double Nickase Plasmid (h): sc-409587-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AGA 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
- AGA Double Nickase Plasmid (h) and AGA Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting AGA. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: AGA Antibody (H-8): sc-514075
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AGA Double Nickase Plasmid (h) | sc-409587-NIC | 20 µg | $410.00 | |||
AGA Double Nickase Plasmid (h2) | sc-409587-NIC-2 | 20 µg | $410.00 |
Human AGA encodes aspartylglucosaminidase, a lysosomal hydrolase that cleaves the N-linked glycoasparagine linkage in glycoprotein catabolism, supporting turnover of glycoprotein-derived glycoamino acids. The enzyme functions within the endo-lysosomal degradation network and contributes to cellular proteostasis by enabling efficient processing of complex glycoconjugates. Disruption of AGA activity is linked to impaired lysosomal clearance and accumulation of undegraded substrates, a hallmark of lysosomal storage biology. Consequently, AGA is frequently studied in the context of lysosome-dependent metabolism, stress responses to proteotoxic burden, and genotype–phenotype relationships in inherited metabolic disease models.
AGA Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the AGA locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within AGA. 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 AGA 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 AGA-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.