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.
HRI Double Nickase Plasmid (h): sc-403442-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HRI 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
- HRI Double Nickase Plasmid (h) and HRI Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EIF2AK1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HRI Antibody (D-12): sc-365239
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HRI Double Nickase Plasmid (h) | sc-403442-NIC | 20 µg | $410.00 |
EIF2AK1 encodes heme-regulated inhibitor (HRI), a stress-sensing eIF2α kinase that phosphorylates EIF2S1 to modulate translation initiation and the integrated stress response. In erythroid and non-erythroid contexts, HRI couples heme availability, oxidative stress, and proteotoxic stress to attenuation of global protein synthesis while promoting adaptive transcriptional programs such as ATF4 signaling. This regulation impacts proteostasis, redox homeostasis, and mitochondrial and cytosolic stress pathways that shape cell survival under nutrient and iron/heme fluctuations. Dysregulated EIF2AK1–eIF2α signaling has been investigated in anemia-related erythropoietic stress, neurodegeneration-associated proteostasis defects, and tumor cell adaptation to microenvironmental stress.
HRI Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EIF2AK1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EIF2AK1. 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 EIF2AK1 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 EIF2AK1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.