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.
Erythropoietin/EPO Double Nickase Plasmid (h): sc-400283-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Erythropoietin/EPO 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
- Erythropoietin/EPO Double Nickase Plasmid (h) and Erythropoietin/EPO Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EPO. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Erythropoietin/EPO Antibody (7D10): sc-80995
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Erythropoietin/EPO Double Nickase Plasmid (h) | sc-400283-NIC | 20 µg | $410.00 | |||
Erythropoietin/EPO Double Nickase Plasmid (h2) | sc-400283-NIC-2 | 20 µg | $410.00 |
Human EPO encodes erythropoietin, a secreted glycoprotein hormone that regulates erythroid progenitor survival, proliferation, and differentiation through engagement of EPOR and downstream JAK2/STAT5 signaling. This pathway integrates with PI3K–AKT and MAPK cascades to coordinate anti-apoptotic programs and erythropoietic output under hypoxic stress, including transcriptional control by HIF factors. Beyond hematopoiesis, EPO/EPOR signaling is used to probe cytokine receptor biology, stress-response transcriptional networks, and secreted factor dynamics in cellular crosstalk. Dysregulated EPO expression or signaling is frequently studied in the context of anemia biology, hypoxia-associated remodeling, and tumor microenvironment adaptation where oxygen-sensing pathways are perturbed.
Erythropoietin/EPO Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EPO locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EPO. 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 EPO 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 EPO-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.