E2F-5 Double Nickase Plasmid (h): sc-401915-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- E2F-5 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
- E2F-5 Double Nickase Plasmid (h) and E2F-5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting E2F5. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: E2F-5 Antibody (C-8): sc-374268
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
E2F-5 Double Nickase Plasmid (h) | sc-401915-NIC | 20 µg | $410.00 | |||
E2F-5 Double Nickase Plasmid (h2) | sc-401915-NIC-2 | 20 µg | $410.00 |
E2F5 encodes E2F-5, a member of the E2F family of transcription factors that coordinates cell-cycle control by regulating genes required for G1/S transition, DNA replication, and checkpoint responses. E2F-5 function is closely linked to the RB/E2F axis, integrating cyclin–CDK activity with transcriptional programs that shape proliferation, differentiation, and quiescence. Through promoter binding in concert with DP proteins and RB family repressors, E2F-5 influences chromatin and transcriptional output in pathways governing growth control and cellular fate decisions. Dysregulated E2F5 signaling has been associated with aberrant proliferative states and tumor-associated transcriptional programs, making it a useful node for studying cell-cycle misregulation in disease-relevant contexts.
E2F-5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the E2F5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within E2F5. 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 E2F5 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 E2F5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.