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.
AFX1 Double Nickase Plasmid (h): sc-402408-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AFX1 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
- AFX1 Double Nickase Plasmid (h) and AFX1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting FOXO4. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: AFX1 Antibody (A-7): sc-373877
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AFX1 Double Nickase Plasmid (h) | sc-402408-NIC | 20 µg | $410.00 | |||
AFX1 Double Nickase Plasmid (h2) | sc-402408-NIC-2 | 20 µg | $410.00 |
FOXO4 (AFX1) is a forkhead box O transcription factor that integrates insulin/IGF-1 signaling, PI3K–AKT phosphorylation dynamics, and stress-responsive pathways to regulate genes controlling cell-cycle arrest, apoptosis, oxidative stress resistance, and metabolism. Its activity is modulated by post-translational modifications that influence nuclear–cytoplasmic shuttling and DNA binding, thereby shaping transcriptional programs linked to cellular homeostasis. FOXO4 participates in network crosstalk with MAPK, AMPK, and TGF-β signaling and can influence chromatin accessibility through cofactor recruitment. Dysregulated FOXO4 signaling has been associated with altered proliferation and stress adaptation in cancer biology, as well as cellular senescence and tissue aging mechanisms.
AFX1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the FOXO4 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within FOXO4. 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 FOXO4 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 FOXO4-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.