Cdc25A Double Nickase Plasmid (h): sc-400345-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Cdc25A 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
- Cdc25A Double Nickase Plasmid (h) and Cdc25A Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CDC25A. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Cdc25A Antibody (F-6): sc-7389
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Cdc25A Double Nickase Plasmid (h) | sc-400345-NIC | 20 µg | $410.00 | |||
Cdc25A Double Nickase Plasmid (h2) | sc-400345-NIC-2 | 20 µg | $410.00 |
CDC25A encodes Cdc25A, a dual-specificity phosphatase that activates cyclin-dependent kinases by removing inhibitory phosphates on CDK2 and CDK1, thereby promoting G1/S transition and S-phase progression. Its activity is tightly controlled by checkpoint signaling, including ATR/CHK1-mediated phosphorylation that triggers ubiquitin-dependent degradation in response to replication stress and DNA damage. Through these pathways, Cdc25A coordinates cell-cycle timing with genome integrity surveillance and interacts functionally with p53- and RB-linked networks. Dysregulated CDC25A expression or stability is frequently associated with aberrant proliferation phenotypes and is studied in the context of oncogenic signaling and checkpoint defects.
Cdc25A Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CDC25A locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CDC25A. 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 CDC25A 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 CDC25A-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.