AGXT Double Nickase Plasmid (h): sc-407697-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AGXT 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
- AGXT Double Nickase Plasmid (h) and AGXT Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting AGXT. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: AGXT Antibody (173J2B): sc-517624
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AGXT Double Nickase Plasmid (h) | sc-407697-NIC | 20 µg | $410.00 | |||
AGXT Double Nickase Plasmid (h2) | sc-407697-NIC-2 | 20 µg | $410.00 |
Human AGXT encodes alanine–glyoxylate aminotransferase, a peroxisomal PLP-dependent enzyme that catalyzes transamination of glyoxylate to glycine using alanine as the amino donor. This activity is central to hepatic glyoxylate detoxification and links amino acid metabolism with peroxisomal homeostasis, limiting conversion of glyoxylate to oxalate. Disruption of AGXT function is associated with primary hyperoxaluria type 1, characterized by elevated oxalate production and calcium oxalate deposition, making AGXT a key node for studying peroxisomal metabolic control. AGXT also serves as a model gene for investigating subcellular targeting, enzyme cofactor dependence, and metabolic stress responses in human cells.
AGXT Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the AGXT locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within AGXT. 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 AGXT 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 AGXT-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.