Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
3PGDH Double Nickase Plasmid (h): sc-401405-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- 3PGDH 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
- 3PGDH Double Nickase Plasmid (h) and 3PGDH Double Nickase Plasmid (h2) 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 (h) | sc-401405-NIC | 20 µg | $410.00 | |||
3PGDH Double Nickase Plasmid (h2) | sc-401405-NIC-2 | 20 µg | $410.00 |
PHGDH encodes 3-phosphoglycerate dehydrogenase (3PGDH), the rate-limiting enzyme that diverts the glycolytic intermediate 3-phosphoglycerate into the phosphorylated serine biosynthesis pathway. By producing 3-phosphohydroxypyruvate and supporting downstream serine and glycine availability, 3PGDH contributes to one-carbon metabolism, nucleotide synthesis, redox balance through NADPH-linked reactions, and biosynthetic flux required for proliferative states. Altered PHGDH activity has been associated with metabolic rewiring in cancer models and with neurodevelopmental disorders linked to serine deficiency, underscoring its relevance to studies of metabolism-driven cellular phenotypes. These functions make PHGDH a useful node for interrogating crosstalk between glycolysis, amino acid homeostasis, and mitochondrial/folate-cycle-dependent processes.
3PGDH Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PHGDH locus in human 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.