E2F1 Double Nickase Plasmid (h): sc-400123-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- E2F1 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
- E2F1 Double Nickase Plasmid (h) and E2F1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting E2F1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: E2F1 Antibody (KH95): sc-251
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
E2F1 Double Nickase Plasmid (h) | sc-400123-NIC | 20 µg | $410.00 | |||
E2F1 Double Nickase Plasmid (h2) | sc-400123-NIC-2 | 20 µg | $410.00 |
E2F1 encodes a sequence-specific transcription factor that integrates mitogenic cues with cell-cycle progression by activating genes required for G1/S transition and DNA replication. E2F1 activity is tightly controlled by the RB1 pathway and cyclin-dependent kinase signaling, and it participates in checkpoints that coordinate proliferation with apoptosis and DNA damage responses. Dysregulated E2F1 signaling is frequently associated with aberrant cell-cycle control and genomic instability, making it relevant to studies of oncogenic transformation, replication stress, and tumor suppressor networks. As a downstream effector of RB/E2F and p53-linked stress pathways, E2F1 is widely used to probe transcriptional control of proliferation and cell fate decisions.
E2F1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the E2F1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within E2F1. 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 E2F1 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 E2F1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.