ACOT8 Double Nickase Plasmid (h): sc-407262-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ACOT8 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
- ACOT8 Double Nickase Plasmid (h) and ACOT8 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ACOT8. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ACOT8 Antibody (C-3): sc-7343
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ACOT8 Double Nickase Plasmid (h) | sc-407262-NIC | 20 µg | $410.00 | |||
ACOT8 Double Nickase Plasmid (h2) | sc-407262-NIC-2 | 20 µg | $410.00 |
ACOT8 encodes acyl-CoA thioesterase 8, a peroxisomal enzyme that hydrolyzes fatty acyl-CoA thioesters to free fatty acids and coenzyme A, helping regulate intraperoxisomal CoA availability and acyl-CoA pool size. By modulating substrate flux through peroxisomal β-oxidation and lipid remodeling, ACOT8 contributes to cellular lipid homeostasis and redox balance. Its activity interfaces with broader peroxisome-associated processes including very-long-chain fatty acid handling and coordination with mitochondrial fatty acid oxidation. Dysregulation of peroxisomal lipid metabolism pathways involving ACOT8 has been investigated in the context of metabolic and inflammatory phenotypes where altered fatty acid utilization and oxidative stress are relevant.
ACOT8 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ACOT8 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ACOT8. 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 ACOT8 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 ACOT8-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.