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.
citrate synthase Double Nickase Plasmid (h): sc-402227-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- citrate synthase 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
- citrate synthase Double Nickase Plasmid (h) and citrate synthase Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CS. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: citrate synthase Antibody (G-3): sc-390693
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
citrate synthase Double Nickase Plasmid (h) | sc-402227-NIC | 20 µg | $410.00 | |||
citrate synthase Double Nickase Plasmid (h2) | sc-402227-NIC-2 | 20 µg | $410.00 |
Human CS encodes citrate synthase, a mitochondrial matrix enzyme that catalyzes the condensation of oxaloacetate and acetyl‑CoA to form citrate, initiating flux through the tricarboxylic acid (TCA) cycle. By controlling entry into oxidative metabolism, citrate synthase supports NADH/FADH2 production for electron transport and influences anaplerosis/cataplerosis linking carbohydrate, lipid, and amino acid metabolism. CS activity intersects with mitochondrial bioenergetics, redox homeostasis, and citrate-dependent biosynthetic processes such as fatty acid and sterol synthesis. Altered mitochondrial metabolism involving TCA cycle function is frequently examined in contexts including metabolic dysregulation, neurodegeneration, and proliferative phenotypes where mitochondrial capacity and substrate utilization are remodeled.
citrate synthase Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CS locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CS. 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 CS 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 CS-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.