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.
PKM Double Nickase Plasmid (h): sc-400834-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PKM 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
- PKM Double Nickase Plasmid (h) and PKM Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PKM. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PKM Antibody (C-11): sc-365684
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PKM Double Nickase Plasmid (h) | sc-400834-NIC | 20 µg | $410.00 | |||
PKM Double Nickase Plasmid (h2) | sc-400834-NIC-2 | 20 µg | $410.00 |
PKM encodes pyruvate kinase M, a rate-limiting glycolytic enzyme that catalyzes phosphoenolpyruvate-to-pyruvate conversion with concomitant ATP generation, thereby coupling glucose catabolism to cellular energy balance. Alternative isoforms (PKM1/PKM2) support distinct metabolic states, with PKM2 enabling diversion of glycolytic intermediates into anabolic pathways that sustain biomass production and redox homeostasis. PKM activity interfaces with central carbon metabolism, including the pentose phosphate pathway and lactate production, shaping nutrient sensing and stress responses. Dysregulated PKM expression or isoform usage is frequently studied in contexts of metabolic reprogramming, proliferation, and altered bioenergetics across diverse disease models.
PKM Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PKM locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PKM. 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 PKM 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 PKM-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.