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.
FKHRL1/FOXO3a Double Nickase Plasmid (h): sc-400308-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- FKHRL1/FOXO3a 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
- FKHRL1/FOXO3a Double Nickase Plasmid (h) and FKHRL1/FOXO3a Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting FOXO3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: FKHRL1/FOXO3a Antibody (D-12): sc-48348
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
FKHRL1/FOXO3a Double Nickase Plasmid (h) | sc-400308-NIC | 20 µg | $410.00 | |||
FKHRL1/FOXO3a Double Nickase Plasmid (h2) | sc-400308-NIC-2 | 20 µg | $410.00 |
FOXO3 (FKHRL1/FOXO3a) encodes a forkhead box O transcription factor that integrates growth factor and nutrient signaling to regulate cell-cycle control, oxidative stress resistance, autophagy, and apoptosis. FOXO3 activity is tightly controlled by post-translational modifications, including AKT-mediated phosphorylation downstream of PI3K–AKT that drives cytoplasmic sequestration, and stress-activated inputs such as AMPK and JNK that promote nuclear localization and transcriptional output. Through regulation of targets involved in metabolism, DNA damage responses, and proteostasis, FOXO3 contributes to cellular homeostasis across multiple lineages. Dysregulated FOXO3 signaling has been associated with altered stress tolerance and survival programs relevant to cancer biology, neurodegeneration, metabolic dysfunction, and immune regulation.
FKHRL1/FOXO3a Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the FOXO3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within FOXO3. 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 FOXO3 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 FOXO3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.