PKR Double Nickase Plasmid (h): sc-400177-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PKR 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
- PKR Double Nickase Plasmid (h) and PKR Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EIF2AK2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PKR Antibody (B-10): sc-6282
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PKR Double Nickase Plasmid (h) | sc-400177-NIC | 20 µg | $410.00 | |||
PKR Double Nickase Plasmid (h2) | sc-400177-NIC-2 | 20 µg | $410.00 |
EIF2AK2 encodes PKR, an interferon-inducible serine/threonine kinase that functions as a cytosolic sensor of double-stranded RNA during antiviral and innate immune responses. Upon activation, PKR phosphorylates eIF2α to suppress translation initiation, reshaping cellular proteostasis and promoting stress-adaptive programs such as the integrated stress response. PKR also intersects with NF-κB and MAPK signaling, contributing to inflammatory gene expression, apoptosis, and autophagy regulation. Dysregulated EIF2AK2/PKR activity has been linked to altered host–pathogen interactions, chronic inflammation, and stress-related signaling changes observed across diverse disease contexts, supporting its utility as a mechanistic node in immunology and cell-stress research.
PKR Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EIF2AK2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EIF2AK2. 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 EIF2AK2 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 EIF2AK2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.