IRE1α Double Nickase Plasmid (h): sc-400576-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- IRE1α 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
- IRE1α Double Nickase Plasmid (h) and IRE1α Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ERN1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: IRE1α Antibody (B-12): sc-390960
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
IRE1α Double Nickase Plasmid (h) | sc-400576-NIC | 20 µg | $410.00 | |||
IRE1α Double Nickase Plasmid (h2) | sc-400576-NIC-2 | 20 µg | $410.00 |
ERN1 encodes inositol-requiring enzyme 1 alpha (IRE1α), an ER-resident transmembrane kinase/endoribonuclease that serves as a primary sensor of unfolded proteins and a central hub of the unfolded protein response (UPR). Upon ER stress, IRE1α oligomerizes and activates RNase-mediated splicing of XBP1 mRNA and regulated IRE1-dependent decay (RIDD), reshaping transcriptional and translational programs that govern proteostasis, secretion capacity, inflammation, and apoptosis. Through crosstalk with PERK and ATF6 signaling and downstream engagement of JNK and NF-κB pathways, ERN1 influences innate immune signaling and metabolic adaptation. Dysregulated IRE1α–XBP1 activity is implicated in conditions characterized by chronic ER stress, including cancer cell survival programs, neurodegeneration, diabetes, and inflammatory disorders, making it a common target for mechanistic studies.
IRE1α Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ERN1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ERN1. 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 ERN1 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 ERN1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.