eIF4E Double Nickase Plasmid (h): sc-400415-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- eIF4E 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
- eIF4E Double Nickase Plasmid (h) and eIF4E Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EIF4E. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: eIF4E Antibody (P-2): sc-9976
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
eIF4E Double Nickase Plasmid (h) | sc-400415-NIC | 20 µg | $410.00 | |||
eIF4E Double Nickase Plasmid (h2) | sc-400415-NIC-2 | 20 µg | $410.00 |
EIF4E encodes eIF4E, the cap-binding subunit of the eIF4F translation initiation complex that recognizes the 7-methylguanosine cap of mRNAs and promotes recruitment of ribosomes to initiate protein synthesis. eIF4E activity integrates signals from PI3K–AKT–mTOR and MAPK pathways through regulation by 4E-BP proteins and MNK-dependent phosphorylation, coupling growth cues to selective translation of transcripts involved in proliferation, survival, and stress responses. Dysregulated EIF4E expression or signaling control is linked to aberrant translational reprogramming in cancer biology and has also been implicated in neurodevelopmental and viral-host interaction studies. As a central node in cap-dependent translation, EIF4E is widely used to interrogate translational control, mRNA selection, and proteostasis-related phenotypes in human cell models.
eIF4E Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EIF4E locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EIF4E. 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 EIF4E 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 EIF4E-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.