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.
creatine kinase-M Double Nickase Plasmid (h): sc-401132-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- creatine kinase-M 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
- creatine kinase-M Double Nickase Plasmid (h) and creatine kinase-M Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CKM. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: creatine kinase-M Antibody (G-9): sc-365046
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
creatine kinase-M Double Nickase Plasmid (h) | sc-401132-NIC | 20 µg | $410.00 | |||
creatine kinase-M Double Nickase Plasmid (h2) | sc-401132-NIC-2 | 20 µg | $410.00 |
CKM encodes the muscle-type creatine kinase (creatine kinase-M), a cytosolic phosphotransferase that catalyzes reversible transfer of phosphate between ATP and creatine to buffer and rapidly regenerate ATP during fluctuating energy demand. This reaction is central to the phosphocreatine shuttle and links glycolysis and oxidative phosphorylation to myofibrillar ATP consumption, supporting muscle contraction and broader cellular energy homeostasis. CKM activity contributes to creatine/phosphagen metabolism and is commonly used as a biochemical and molecular readout in studies of skeletal muscle differentiation, stress responses, and mitochondrial function. Dysregulated creatine kinase signaling and altered CKM expression have been associated with muscle damage and metabolic perturbations, making it relevant for mechanistic investigations of myopathies and related energy-deficit phenotypes.
creatine kinase-M Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CKM locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CKM. 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 CKM 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 CKM-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.