c-Kit Double Nickase Plasmid (m): sc-421289-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- c-Kit Double Nickase Plasmid (m) 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
- c-Kit Double Nickase Plasmid (m) and c-Kit Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Kit. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: c-Kit Antibody (E-3): sc-365504
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
c-Kit Double Nickase Plasmid (m) | sc-421289-NIC | 20 µg | $410.00 | |||
c-Kit Double Nickase Plasmid (m2) | sc-421289-NIC-2 | 20 µg | $410.00 |
Kit (c-Kit; Cd117) encodes a receptor tyrosine kinase for stem cell factor that regulates survival, proliferation, and lineage commitment in hematopoietic progenitors, melanocytes, germ cells, and mast cells. Upon ligand-induced dimerization and autophosphorylation, c-Kit engages PI3K–AKT, RAS–MAPK, JAK/STAT, and SRC-family signaling to coordinate cell-cycle progression, migration, and differentiation. Dysregulated KIT signaling and KIT-expressing cell populations are widely studied in tumor biology, mast cell and allergy-related inflammation, and hematopoietic developmental defects, supporting its use as a pathway node for mechanistic studies. In mouse systems, Kit function is also leveraged to interrogate stem/progenitor cell dynamics and microenvironmental cues in development and tissue maintenance.
c-Kit Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Kit locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Kit. 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 Kit 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 Kit-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.