TRIM5α Double Nickase Plasmid (h): sc-409348-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TRIM5α 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
- TRIM5α Double Nickase Plasmid (h) and TRIM5α Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting TRIM5. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TRIM5α Double Nickase Plasmid (h) | sc-409348-NIC | 20 µg | $410.00 | |||
TRIM5α Double Nickase Plasmid (h2) | sc-409348-NIC-2 | 20 µg | $410.00 |
TRIM5 (TRIM5α) is an interferon-inducible tripartite motif E3 ubiquitin ligase that acts as an intrinsic restriction factor against retroviral infection by recognizing incoming capsids and triggering premature uncoating. Through its RING-dependent ubiquitination activity and higher-order lattice assembly, TRIM5α engages innate immune signaling, including NF-κB and AP-1 pathways, and interfaces with proteasome- and autophagy-linked protein quality control. TRIM5α also participates in cytosolic pattern recognition and contributes to shaping inflammatory transcriptional programs following viral challenge. Variation or altered regulation of TRIM5 has been studied in the context of susceptibility to retroviral replication, antiviral immune responsiveness, and broader dysregulation of innate immune signaling relevant to inflammatory disease models.
TRIM5α Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the TRIM5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within TRIM5. 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 TRIM5 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 TRIM5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.