TRAC-1 Double Nickase Plasmid (h): sc-409980-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TRAC-1 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
- TRAC-1 Double Nickase Plasmid (h) and TRAC-1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RNF125. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TRAC-1 Double Nickase Plasmid (h) | sc-409980-NIC | 20 µg | $410.00 | |||
TRAC-1 Double Nickase Plasmid (h2) | sc-409980-NIC-2 | 20 µg | $410.00 |
RNF125 encodes the human E3 ubiquitin ligase TRAC-1, a RING finger protein that helps control protein turnover and signal amplitude in immune cells through ubiquitin-dependent proteasomal pathways. TRAC-1 has been implicated in regulation of T cell receptor and cytokine signaling outputs by modulating stability and trafficking of signaling components, thereby shaping activation, tolerance, and downstream transcriptional programs. As part of broader ubiquitination networks, RNF125 intersects with innate and adaptive immune regulatory circuits that influence inflammatory responses and antigen-driven lymphocyte function. Dysregulated RNF125/TRAC-1 activity and altered expression patterns have been associated with immune-mediated and inflammatory disease mechanisms, supporting its use as a target for studying immune homeostasis and signaling rewiring.
TRAC-1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RNF125 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RNF125. 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 RNF125 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 RNF125-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.