eRF1 Double Nickase Plasmid (h): sc-405156-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- eRF1 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
- eRF1 Double Nickase Plasmid (h) and eRF1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ETF1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: eRF1 Antibody (B-11): sc-365686
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
eRF1 Double Nickase Plasmid (h) | sc-405156-NIC | 20 µg | $410.00 | |||
eRF1 Double Nickase Plasmid (h2) | sc-405156-NIC-2 | 20 µg | $410.00 |
Human ETF1 encodes eukaryotic release factor 1 (eRF1), an essential translation termination factor that recognizes all three stop codons and promotes peptidyl-tRNA hydrolysis at the ribosomal A site in concert with eRF3/GSPT1. Through its central role in proteostasis, eRF1 helps maintain fidelity of mRNA translation and limits stop-codon readthrough, linking ETF1 activity to ribosome quality control and surveillance pathways such as nonsense-mediated mRNA decay. Perturbation of translation termination can reshape stress-response signaling, protein folding burden, and global gene expression programs relevant to cellular homeostasis. ETF1 is therefore of interest in studies of translational control, codon recognition, and mechanisms that couple ribosome dynamics to disease-associated proteome imbalance.
eRF1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ETF1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ETF1. 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 ETF1 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 ETF1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.