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.
EED Double Nickase Plasmid (h): sc-401234-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- EED 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
- EED Double Nickase Plasmid (h) and EED Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EED. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: EED Antibody (3B12): sc-293203
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
EED Double Nickase Plasmid (h) | sc-401234-NIC | 20 µg | $410.00 | |||
EED Double Nickase Plasmid (h2) | sc-401234-NIC-2 | 20 µg | $410.00 |
EED encodes a core component of the Polycomb Repressive Complex 2 (PRC2) that binds trimethylated histone H3 lysine 27 (H3K27me3) and allosterically stimulates EZH2/EZH1 methyltransferase activity to propagate repressive chromatin domains. Through PRC2-dependent gene silencing, EED contributes to maintenance of cell identity, control of developmental transcriptional programs, and regulation of proliferation and differentiation. Perturbation of EED disrupts epigenetic repression and can alter pathways governing lineage commitment and chromatin organization. Dysregulated PRC2/EED function is implicated across cancers and developmental disorders, making EED a common target in studies of epigenetic control and transcriptional repression.
EED Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EED locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EED. 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 EED 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 EED-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.