3PGDH Double Nickase Plasmid (m): sc-433506-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- 3PGDH Double Nickase Plasmid (m) 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
- 3PGDH Double Nickase Plasmid (m) and 3PGDH Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Phgdh. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: 3PGDH Antibody (B-1): sc-390610
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
3PGDH Double Nickase Plasmid (m) | sc-433506-NIC | 20 µg | $410.00 | |||
3PGDH Double Nickase Plasmid (m2) | sc-433506-NIC-2 | 20 µg | $410.00 |
Mouse Phgdh encodes 3-phosphoglycerate dehydrogenase (3PGDH), the first and rate-limiting enzyme of the phosphorylated L-serine biosynthesis pathway that diverts the glycolytic intermediate 3-phosphoglycerate toward serine and glycine production. Through control of serine availability, 3PGDH influences one-carbon metabolism, nucleotide synthesis, redox homeostasis, and lipid biosynthetic programs that support proliferation and differentiation. Phgdh activity is therefore closely linked to metabolic reprogramming and cellular stress responses in tissues with high biosynthetic demand, including the nervous system. Altered PHGDH/3PGDH function has been associated with inborn errors of serine metabolism and with metabolic dependencies observed in some proliferative states, making Phgdh a useful node for studying disease-relevant metabolic circuitry.
3PGDH Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Phgdh locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Phgdh. 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 Phgdh 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 Phgdh-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.