β Enolase Double Nickase Plasmid (h): sc-400843-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- β Enolase 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
- β Enolase Double Nickase Plasmid (h) and β Enolase Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ENO3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: β Enolase Antibody (XX-10): sc-100811
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
β Enolase Double Nickase Plasmid (h) | sc-400843-NIC | 20 µg | $410.00 | |||
β Enolase Double Nickase Plasmid (h2) | sc-400843-NIC-2 | 20 µg | $410.00 |
ENO3 encodes human β enolase (ENO3), a muscle-enriched glycolytic enzyme that catalyzes the conversion of 2-phosphoglycerate to phosphoenolpyruvate, supporting ATP production in high-energy tissues. Beyond glycolysis, enolase isoforms contribute to metabolic rewiring and cellular stress responses through links to hypoxia signaling and broader carbon flux regulation. Altered ENO3 expression or activity has been associated with skeletal muscle pathology and myopathic phenotypes, and is studied in contexts of metabolic dysfunction and tissue remodeling. As a marker of muscle metabolic state, ENO3 is frequently used to interrogate changes in energy metabolism, differentiation, and proteostasis in human cell models.
β Enolase Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ENO3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ENO3. 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 ENO3 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 ENO3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.