eEF2K Double Nickase Plasmid (h): sc-402166-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- eEF2K 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
- eEF2K Double Nickase Plasmid (h) and eEF2K Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EEF2K. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: eEF2K Antibody (C-12): sc-390710
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
eEF2K Double Nickase Plasmid (h) | sc-402166-NIC | 20 µg | $410.00 | |||
eEF2K Double Nickase Plasmid (h2) | sc-402166-NIC-2 | 20 µg | $410.00 |
EEF2K encodes eukaryotic elongation factor 2 kinase (eEF2K), a Ca2+/calmodulin-dependent Ser/Thr kinase that phosphorylates eEF2 to slow ribosomal translocation and restrain protein synthesis during cellular stress. Acting downstream of nutrient and energy sensing networks including mTOR and AMPK signaling, eEF2K helps coordinate translational control with autophagy and metabolic adaptation. This pathway influences cell growth, survival, and synaptic plasticity by tuning global and selective mRNA translation. Altered eEF2K activity and eEF2 phosphorylation have been reported in cancer-associated stress responses and in neurological and metabolic disorders, supporting its relevance for mechanistic studies.
eEF2K Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EEF2K locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EEF2K. 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 EEF2K 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 EEF2K-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.