eIF2α Double Nickase Plasmid (h): sc-400199-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- eIF2α 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
- eIF2α Double Nickase Plasmid (h) and eIF2α Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EIF2S1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: eIF2α Antibody (D-3): sc-133132
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
eIF2α Double Nickase Plasmid (h) | sc-400199-NIC | 20 µg | $410.00 | |||
eIF2α Double Nickase Plasmid (h2) | sc-400199-NIC-2 | 20 µg | $410.00 |
EIF2S1 encodes the eukaryotic translation initiation factor 2 alpha subunit (eIF2α), a central regulator of cap-dependent translation initiation and a key node in the integrated stress response. Phosphorylation of eIF2α at Ser51 by stress-responsive kinases such as PERK (EIF2AK3), GCN2 (EIF2AK4), PKR (EIF2AK2), and HRI (EIF2AK1) reduces ternary complex availability, globally attenuating protein synthesis while enabling selective translation programs. This signaling axis coordinates proteostasis, ER stress adaptation, autophagy, and innate immune responses through downstream effectors including ATF4 and CHOP/DDIT3. Dysregulated eIF2α signaling is implicated in contexts such as neurodegeneration, cancer cell stress tolerance, viral infection responses, and metabolic stress, making EIF2S1 a widely used handle for probing translational control mechanisms.
eIF2α Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EIF2S1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EIF2S1. 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 EIF2S1 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 EIF2S1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.