eIF2A Double Nickase Plasmid (h): sc-413527-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- eIF2A 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
- eIF2A Double Nickase Plasmid (h) and eIF2A Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EIF2A. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: eIF2A Antibody (3A7A8): sc-517214
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
eIF2A Double Nickase Plasmid (h) | sc-413527-NIC | 20 µg | $410.00 |
EIF2A encodes eIF2A, a non-canonical translation initiation factor that can deliver initiator Met-tRNAi to the 40S ribosomal subunit independently of the eIF2-GTP ternary complex. eIF2A activity is particularly relevant during cellular stress conditions that attenuate canonical initiation, linking EIF2A to translational reprogramming and proteostasis control. Through its role in regulating mRNA-specific initiation and stress-adaptive protein synthesis, EIF2A intersects with integrated stress response and related signaling nodes that influence cell survival and homeostasis. Dysregulated translation control involving EIF2A has been investigated in contexts such as tumor cell stress adaptation and neurodegeneration-associated proteotoxic stress, supporting its utility as a mechanistic target in pathway studies.
eIF2A Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EIF2A locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EIF2A. 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 EIF2A 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 EIF2A-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.