CAT Double Nickase Plasmid (h): sc-404860-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CAT 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
- CAT Double Nickase Plasmid (h) and CAT Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CRAT. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CAT Antibody (654D2Y): sc-517650
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CAT Double Nickase Plasmid (h) | sc-404860-NIC | 20 µg | $410.00 | |||
CAT Double Nickase Plasmid (h2) | sc-404860-NIC-2 | 20 µg | $410.00 |
CRAT encodes carnitine O-acetyltransferase (CAT), a mitochondrial matrix enzyme that catalyzes reversible transfer of acetyl groups between acetyl-CoA and carnitine to form acetylcarnitine. This reaction links fatty acid β-oxidation, pyruvate-derived acetyl-CoA utilization, and maintenance of mitochondrial CoA pools, thereby influencing redox balance and metabolic flexibility. By modulating acetyl group buffering and acylcarnitine profiles, CAT contributes to regulation of energy metabolism during nutrient shifts and stress. Dysregulated carnitine-dependent acetyl flux and acylcarnitine accumulation are widely used biochemical readouts in studies of metabolic syndrome, insulin resistance, and mitochondrial dysfunction, making CRAT a useful node for mechanistic interrogation.
CAT Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CRAT locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CRAT. 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 CRAT 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 CRAT-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.