Oct3/4 Double Nickase Plasmid (h): sc-410951-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Oct3/4 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
- Oct3/4 Double Nickase Plasmid (h) and Oct3/4 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting POU5F1B. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Oct3/4 Antibody (A-9): sc-365509
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Oct3/4 Double Nickase Plasmid (h) | sc-410951-NIC | 20 µg | $410.00 | |||
Oct3/4 Double Nickase Plasmid (h2) | sc-410951-NIC-2 | 20 µg | $410.00 |
POU5F1B encodes a human Oct3/4-related POU domain transcription factor implicated in the regulation of pluripotency-associated transcriptional programs and chromatin state. Oct3/4 family activity interfaces with core stemness networks and transcriptional circuitry governing self-renewal, lineage commitment, and reprogramming, with downstream effects on cell cycle control and epigenetic remodeling. Aberrant expression or dysregulation of POU5F1B has been reported in multiple tumor contexts and is studied in relation to oncogenic transcriptional programs, cellular plasticity, and stress-adaptive phenotypes. This gene is therefore relevant for dissecting mechanisms of stem-like states, differentiation barriers, and transcription factor–driven disease biology in human cell models.
Oct3/4 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the POU5F1B locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within POU5F1B. 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 POU5F1B 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 POU5F1B-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.