eRF3a Double Nickase Plasmid (h): sc-404748-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- eRF3a 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
- eRF3a Double Nickase Plasmid (h) and eRF3a Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GSPT1. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
eRF3a Double Nickase Plasmid (h) | sc-404748-NIC | 20 µg | $410.00 | |||
eRF3a Double Nickase Plasmid (h2) | sc-404748-NIC-2 | 20 µg | $410.00 |
Human GSPT1 encodes eRF3a, a translation termination factor that forms a functional complex with eRF1 to promote GTP-dependent peptide release at stop codons and coordinate ribosome recycling. Beyond termination, eRF3a participates in mRNA surveillance pathways such as nonsense-mediated mRNA decay and influences proteostasis by linking translation dynamics to protein quality control. GSPT1 activity is integrated with cell-cycle regulation and stress-responsive translational programs, and altered expression has been associated with proliferative phenotypes in multiple cancer-relevant contexts. As a core node in translational fidelity and mRNA turnover, eRF3a is frequently studied to interrogate mechanisms of stop-codon recognition, transcript stability, and growth control.
eRF3a Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GSPT1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GSPT1. 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 GSPT1 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 GSPT1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.