MUT Double Nickase Plasmid (h): sc-406555-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MUT 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
- MUT Double Nickase Plasmid (h) and MUT Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting MUT. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: MUT Antibody (D-1): sc-390978
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MUT Double Nickase Plasmid (h) | sc-406555-NIC | 20 µg | $410.00 | |||
MUT Double Nickase Plasmid (h2) | sc-406555-NIC-2 | 20 µg | $410.00 |
Human MUT encodes methylmalonyl‑CoA mutase, a mitochondrial adenosylcobalamin-dependent enzyme that catalyzes the isomerization of L‑methylmalonyl‑CoA to succinyl‑CoA, linking propionate catabolism to the tricarboxylic acid cycle. This activity integrates branched-chain amino acid and odd-chain fatty acid metabolism with mitochondrial energy production and anaplerotic flux. Disruption of MUT function perturbs mitochondrial metabolic homeostasis and is strongly associated with methylmalonic acidemia, a prototypical inborn error of metabolism characterized by methylmalonate accumulation and downstream cellular stress responses. As a result, MUT is widely studied in pathways involving vitamin B12 cofactor utilization, mitochondrial metabolism, and metabolic disease modeling in human cell systems.
MUT Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the MUT locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within MUT. 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 MUT 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 MUT-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.