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.
ICAD Double Nickase Plasmid (h): sc-403560-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ICAD 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
- ICAD Double Nickase Plasmid (h) and ICAD Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting DFFA. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ICAD Antibody (F-8): sc-17816
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ICAD Double Nickase Plasmid (h) | sc-403560-NIC | 20 µg | $410.00 | |||
ICAD Double Nickase Plasmid (h2) | sc-403560-NIC-2 | 20 µg | $410.00 |
DFFA encodes DNA fragmentation factor subunit alpha (ICAD), a critical inhibitor and chaperone for the nuclease DFFB/CAD that governs apoptotic DNA fragmentation. During apoptosis, caspase-3 cleavage of ICAD releases active CAD to generate internucleosomal DNA breaks, linking DFFA to caspase signaling, chromatin condensation, and the execution phase of programmed cell death. ICAD also supports proper folding and stabilization of CAD, making DFFA important for regulated nuclease activity and genome integrity under stress. Dysregulation of DFFA-associated apoptotic pathways has been implicated in altered cell survival and DNA damage responses observed across multiple disease contexts, including cancer and neurodegeneration.
ICAD Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the DFFA locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within DFFA. 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 DFFA 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 DFFA-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.