p14ARF/p16 Double Nickase Plasmid (h): sc-400018-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- p14ARF/p16 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
- p14ARF/p16 Double Nickase Plasmid (h) and p14ARF/p16 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CDKN2A. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: p14ARF Antibody (ARF 4C6/4): sc-53392
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
p14ARF/p16 Double Nickase Plasmid (h) | sc-400018-NIC | 20 µg | $410.00 | |||
p14ARF/p16 Double Nickase Plasmid (h2) | sc-400018-NIC-2 | 20 µg | $410.00 |
CDKN2A encodes two tumor suppressor proteins, p16INK4A and p14ARF, that regulate cell-cycle progression and checkpoint control through distinct pathways. p16 inhibits CDK4/6 to restrain RB phosphorylation and enforce the G1–S transition, while p14ARF antagonizes MDM2 to stabilize p53 and promote cell-cycle arrest or senescence in response to oncogenic stress. Through these RB and p53 network connections, CDKN2A integrates proliferative signaling with genome surveillance and is frequently altered in studies of malignant transformation. Loss or silencing of CDKN2A is widely investigated in contexts involving dysregulated proliferation, evasion of senescence, and altered stress-response signaling.
p14ARF/p16 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CDKN2A locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CDKN2A. 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 CDKN2A 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 CDKN2A-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.