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.
SCF Double Nickase Plasmid (h): sc-416442-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- SCF 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
- SCF Double Nickase Plasmid (h) and SCF Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting KITLG. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: SCF Antibody (G-3): sc-13126
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
SCF Double Nickase Plasmid (h) | sc-416442-NIC | 20 µg | $410.00 | |||
SCF Double Nickase Plasmid (h2) | sc-416442-NIC-2 | 20 µg | $410.00 |
Human KITLG encodes stem cell factor (SCF), a key ligand for the receptor tyrosine kinase KIT (CD117) that regulates survival, proliferation, migration, and differentiation in hematopoietic progenitors, melanocytes, germ cells, and mast cells. SCF–KIT signaling activates downstream PI3K–AKT, RAS–MAPK, and JAK–STAT pathways, coordinating stem cell maintenance and lineage commitment within specialized tissue niches. Dysregulated KITLG/KIT axis activity is implicated in altered pigmentation biology, infertility-related developmental defects, and KIT-driven hematologic and solid tumor contexts, making it a focal node for studying growth factor–dependent signaling and microenvironmental control of cell fate.
SCF Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the KITLG locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within KITLG. 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 KITLG 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 KITLG-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.