Ataxin-3 Double Nickase Plasmid (h): sc-417498-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Ataxin-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
- Ataxin-3 Double Nickase Plasmid (h) and Ataxin-3 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ATXN3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Ataxin-3 Antibody (A-7): sc-398114
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Ataxin-3 Double Nickase Plasmid (h) | sc-417498-NIC | 20 µg | $410.00 |
ATXN3 encodes ataxin-3, a deubiquitinating enzyme that edits ubiquitin chains through its Josephin catalytic domain and multiple ubiquitin-interacting motifs, thereby influencing ubiquitin–proteasome system flux and protein quality control. Ataxin-3 participates in proteostasis networks linked to ER-associated degradation, stress responses, and turnover of misfolded or aggregation-prone substrates, with downstream effects on transcriptional regulation and cytoskeletal organization. Polyglutamine expansion in ATXN3 drives toxic gain-of-function and protein aggregation associated with spinocerebellar ataxia type 3 (Machado–Joseph disease), making ATXN3 a widely used locus for studying neurodegeneration-relevant pathways. In cell models, perturbing ATXN3 supports mechanistic interrogation of ubiquitin signaling, aggregate handling, and neuronal vulnerability phenotypes.
Ataxin-3 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ATXN3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ATXN3. 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 ATXN3 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 ATXN3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.