Pellino 3 Double Nickase Plasmid (h): sc-405232-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Pellino 3 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
- Pellino 3 Double Nickase Plasmid (h) and Pellino 3 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PELI3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Pellino 3 Antibody (B-3): sc-376466
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Pellino 3 Double Nickase Plasmid (h) | sc-405232-NIC | 20 µg | $410.00 |
Human PELI3 encodes Pellino 3, a RING-type E3 ubiquitin ligase that functions as a scaffold and ubiquitination regulator in innate immune signaling. Pellino 3 participates in Toll-like receptor and interleukin-1 receptor pathways by modulating ubiquitin-dependent assembly of signaling complexes that influence NF-κB and MAPK activation and downstream inflammatory gene expression. Through control of protein turnover and signaling amplitude, PELI3 has been studied in the context of host defense, cytokine responses, and cellular stress signaling. Dysregulated pathway activity involving Pellino family ubiquitin ligases is relevant to inflammation-associated disease mechanisms and immune cell functional phenotypes.
Pellino 3 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PELI3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PELI3. 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 PELI3 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 PELI3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.