4E-BP1 Double Nickase Plasmid (h): sc-400398-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- 4E-BP1 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
- 4E-BP1 Double Nickase Plasmid (h) and 4E-BP1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EIF4EBP1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: 4E-BP1 Antibody (P-1): sc-9977
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
4E-BP1 Double Nickase Plasmid (h) | sc-400398-NIC | 20 µg | $410.00 | |||
4E-BP1 Double Nickase Plasmid (h2) | sc-400398-NIC-2 | 20 µg | $410.00 |
EIF4EBP1 encodes 4E-BP1, a phosphorylation-regulated inhibitor of cap-dependent translation that binds EIF4E and restrains assembly of the EIF4F initiation complex. mTORC1-mediated phosphorylation of 4E-BP1 releases EIF4E to promote protein synthesis, linking nutrient and growth factor sensing to translational control, cell growth, and metabolic adaptation. This node integrates PI3K–AKT–mTOR signaling with stress-responsive programs and influences expression of oncogenic and pro-survival mRNAs. Altered 4E-BP1 abundance or phosphorylation state is frequently associated with dysregulated proliferation and metabolism in cancer biology and other mTOR pathway–related disorders.
4E-BP1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EIF4EBP1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EIF4EBP1. 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 EIF4EBP1 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 EIF4EBP1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.