Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
FAT10 Double Nickase Plasmid (h): sc-402682-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- FAT10 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
- FAT10 Double Nickase Plasmid (h) and FAT10 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting UBD. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: FAT10 Antibody (A-8): sc-393630
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
FAT10 Double Nickase Plasmid (h) | sc-402682-NIC | 20 µg | $410.00 |
UBD encodes FAT10 (also known as ubiquitin D), a ubiquitin-like modifier that is rapidly induced by pro-inflammatory cytokines and conjugated to substrate proteins to direct their degradation by the 26S proteasome. FAT10 participates in immunoproteasome regulation, antigen processing, and stress-responsive proteostasis, intersecting with NF-κB signaling and other inflammation-associated pathways. Altered UBD/FAT10 expression has been reported in contexts of chronic inflammation and dysregulated protein turnover, linking it to cellular phenotypes such as altered cell cycle control, apoptosis sensitivity, and metabolic remodeling. These features make UBD a useful node for studying cytokine-driven changes in proteome stability and immune-related signaling programs.
FAT10 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the UBD locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within UBD. 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 UBD 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 UBD-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.