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.
PEPCK-M/PCK2 Double Nickase Plasmid (h): sc-400725-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PEPCK-M/PCK2 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
- PEPCK-M/PCK2 Double Nickase Plasmid (h) and PEPCK-M/PCK2 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PCK2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PEPCK-M/PCK2 Antibody (62-W): sc-130388
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PEPCK-M/PCK2 Double Nickase Plasmid (h) | sc-400725-NIC | 20 µg | $410.00 | |||
PEPCK-M/PCK2 Double Nickase Plasmid (h2) | sc-400725-NIC-2 | 20 µg | $410.00 |
Human PCK2 encodes the mitochondrial phosphoenolpyruvate carboxykinase isoform (PEPCK-M), a key gluconeogenic and anaplerotic enzyme that converts oxaloacetate to phosphoenolpyruvate, linking TCA cycle intermediates to cytosolic biosynthetic pathways. By modulating carbon flux between mitochondrial and cytosolic compartments, PCK2 influences glucose and lipid metabolism, redox balance, and amino acid utilization under nutrient stress. PCK2 activity intersects with pathways governing metabolic adaptation, including gluconeogenesis, cataplerosis, and serine/glycine and glyceroneogenic networks. Dysregulated PCK2 expression or function has been associated with altered metabolic states observed in cancer metabolism, insulin resistance, and other disorders characterized by mitochondrial and glucose homeostasis perturbations.
PEPCK-M/PCK2 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PCK2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PCK2. 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 PCK2 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 PCK2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.