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.
Apaf-1 Double Nickase Plasmid (h): sc-401291-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Apaf-1 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
- Apaf-1 Double Nickase Plasmid (h) and Apaf-1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting APAF1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Apaf-1 Antibody (5E11): sc-65891
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Apaf-1 Double Nickase Plasmid (h) | sc-401291-NIC | 20 µg | $410.00 | |||
Apaf-1 Double Nickase Plasmid (h2) | sc-401291-NIC-2 | 20 µg | $410.00 |
APAF1 encodes apoptotic protease activating factor 1 (Apaf-1), a cytosolic scaffold that initiates the intrinsic mitochondrial apoptosis pathway by binding cytochrome c and dATP to assemble the apoptosome. This complex recruits and activates initiator caspase-9, leading to downstream caspase-3/7 activation, DNA fragmentation, and controlled cell dismantling. Apaf-1 integrates stress signals from mitochondrial outer membrane permeabilization and intersects with BCL-2 family regulation and p53-associated damage responses. Dysregulated APAF1 expression or function has been linked to altered apoptotic thresholds in cancer biology and to cell-loss phenotypes relevant to neurodegeneration and developmental disorders.
Apaf-1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the APAF1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within APAF1. 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 APAF1 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 APAF1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.