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.
CAD Double Nickase Plasmid (h): sc-403028-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CAD 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
- CAD Double Nickase Plasmid (h) and CAD Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting DFFB. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CAD Antibody (F-11): sc-374067
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CAD Double Nickase Plasmid (h) | sc-403028-NIC | 20 µg | $410.00 | |||
CAD Double Nickase Plasmid (h2) | sc-403028-NIC-2 | 20 µg | $410.00 |
DFFB encodes CAD (caspase-activated DNase), the endonuclease responsible for internucleosomal DNA fragmentation during apoptosis. In healthy cells, CAD activity is restrained by its inhibitor ICAD (DFFA) until caspase-dependent cleavage of ICAD releases CAD to cleave chromatin, linking apoptotic signaling to nuclear dismantling. This process intersects with caspase cascade pathways, chromatin organization, and clearance of dying cells, and is commonly used as a molecular readout of programmed cell death. Altered regulation of CAD-mediated DNA fragmentation has been associated with aberrant apoptotic phenotypes and genome integrity defects, supporting its relevance in studies of cancer biology, immune homeostasis, and neurodegeneration.
CAD Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the DFFB locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within DFFB. 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 DFFB 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 DFFB-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.