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.
GCN2 Double Nickase Plasmid (h): sc-402313-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GCN2 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
- GCN2 Double Nickase Plasmid (h) and GCN2 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EIF2AK4. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GCN2 Antibody (F-7): sc-374609
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GCN2 Double Nickase Plasmid (h) | sc-402313-NIC | 20 µg | $410.00 | |||
GCN2 Double Nickase Plasmid (h2) | sc-402313-NIC-2 | 20 µg | $410.00 |
Human EIF2AK4 encodes GCN2, a serine/threonine kinase that senses amino acid limitation through uncharged tRNA binding and phosphorylates eIF2α to initiate the integrated stress response. This signaling reduces global translation while promoting selective translation of stress-adaptive transcripts, coupling nutrient availability to proteostasis, autophagy, and metabolic reprogramming. GCN2 integrates inputs from amino acid starvation and other cellular stresses to modulate ATF4-dependent transcriptional programs and crosstalk with mTORC1 and inflammatory signaling. Dysregulated EIF2AK4/GCN2 activity has been linked in the literature to altered stress tolerance, immune cell function, and metabolic imbalance, making it a relevant node for mechanistic studies of stress-adaptation pathways.
GCN2 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EIF2AK4 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EIF2AK4. 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 EIF2AK4 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 EIF2AK4-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.