Waf1/Cip1/CDKN1A p21 Double Nickase Plasmid (h): sc-400013-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Waf1/Cip1/CDKN1A p21 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
- Waf1/Cip1/CDKN1A p21 Double Nickase Plasmid (h) and Waf1/Cip1/CDKN1A p21 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CDKN1A. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Waf1/Cip1/CDKN1A p21 Antibody (F-5): sc-6246
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Waf1/Cip1/CDKN1A p21 Double Nickase Plasmid (h) | sc-400013-NIC | 20 µg | $410.00 | |||
Waf1/Cip1/CDKN1A p21 Double Nickase Plasmid (h2) | sc-400013-NIC-2 | 20 µg | $410.00 |
CDKN1A encodes p21 (Waf1/Cip1), a cyclin-dependent kinase inhibitor that integrates stress signals to restrain cell-cycle progression at the G1/S checkpoint. Induced prominently by p53 in response to DNA damage, p21 inhibits CDK2- and CDK1-associated complexes and modulates PCNA-dependent DNA replication and repair. Through these interactions, p21 influences senescence, transient cell-cycle arrest, and cell fate decisions during genotoxic stress. Dysregulated CDKN1A/p21 signaling is frequently implicated in oncogenic transformation and tumor suppressor pathway disruption, and it is also relevant to processes such as replication stress tolerance and therapy-induced senescence models.
Waf1/Cip1/CDKN1A p21 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CDKN1A locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CDKN1A. 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 CDKN1A 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 CDKN1A-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.