Rhodanese Double Nickase Plasmid (h): sc-405813-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Rhodanese 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
- Rhodanese Double Nickase Plasmid (h) and Rhodanese Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting TST. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Rhodanese Antibody (G-7): sc-271883
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Rhodanese Double Nickase Plasmid (h) | sc-405813-NIC | 20 µg | $410.00 | |||
Rhodanese Double Nickase Plasmid (h2) | sc-405813-NIC-2 | 20 µg | $410.00 |
Human TST encodes rhodanese (thiosulfate sulfurtransferase), a mitochondrial sulfurtransferase that catalyzes transfer of sulfane sulfur from thiosulfate to cyanide, generating thiocyanate and contributing to cellular detoxification. Rhodanese also participates in sulfur trafficking linked to iron–sulfur cluster homeostasis and redox balance, processes that intersect with mitochondrial metabolism and oxidative stress responses. Altered TST activity has been associated with mitochondrial dysfunction signatures and metabolic phenotypes in tissues with high oxidative capacity, supporting its relevance in studies of stress adaptation and bioenergetic regulation. As a mitochondrial enzyme, rhodanese provides a tractable node to interrogate how sulfur metabolism modulates proteome sulfhydration and downstream signaling.
Rhodanese Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the TST locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within TST. 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 TST 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 TST-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.