Cdc25B Double Nickase Plasmid (h): sc-401457-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Cdc25B 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
- Cdc25B Double Nickase Plasmid (h) and Cdc25B Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CDC25B. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Cdc25B Antibody (DCS-162): sc-56266
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Cdc25B Double Nickase Plasmid (h) | sc-401457-NIC | 20 µg | $410.00 | |||
Cdc25B Double Nickase Plasmid (h2) | sc-401457-NIC-2 | 20 µg | $410.00 |
CDC25B encodes the dual-specificity phosphatase Cdc25B, a key activator of cyclin-dependent kinases that promotes cell-cycle progression by removing inhibitory phosphates from CDK1 and CDK2. By coordinating the G2/M transition and integrating checkpoint cues downstream of DNA damage signaling pathways such as ATM/ATR, Cdc25B helps regulate mitotic entry and replication stress responses. Its activity is controlled by phosphorylation-dependent localization and protein stability, linking it to MAPK signaling and proteasomal degradation. Dysregulated CDC25B expression or activity is associated with aberrant proliferation, chromosomal instability, and oncogenic cell-cycle deregulation, making it a common focus in studies of tumor biology.
Cdc25B Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CDC25B locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CDC25B. 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 CDC25B 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 CDC25B-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.