E2F-4 Double Nickase Plasmid (m): sc-430537-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- E2F-4 Double Nickase Plasmid (m) 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
- E2F-4 Double Nickase Plasmid (m) and E2F-4 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting E2f4. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: E2F-4 Antibody (D-7): sc-398543
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
E2F-4 Double Nickase Plasmid (m) | sc-430537-NIC | 20 µg | $410.00 |
E2F-4 (E2f4) is a transcription factor in the E2F family that regulates cell-cycle progression and cell fate decisions by controlling expression of genes required for G1/S transition and DNA replication. In mouse cells, E2F-4 commonly functions with DP partners and associates with pocket proteins such as p107 and p130 to enforce transcriptional repression during quiescence and differentiation. Through these complexes, E2F-4 contributes to RB/E2F network control of proliferation, checkpoint integrity, and chromatin-linked regulation of gene expression. Dysregulation of E2F pathway signaling is broadly relevant to oncogenic proliferation and altered differentiation states, making E2f4 a useful target for mechanistic studies in cancer biology and developmental models.
E2F-4 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the E2f4 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within E2f4. 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 E2f4 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 E2f4-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.