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.
mitoNEET Double Nickase Plasmid (m): sc-424640-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- mitoNEET Double Nickase Plasmid (m) 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
- mitoNEET Double Nickase Plasmid (m) and mitoNEET Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Cisd1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: mitoNEET Antibody (AT1A8): sc-517413
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
mitoNEET Double Nickase Plasmid (m) | sc-424640-NIC | 20 µg | $410.00 | |||
mitoNEET Double Nickase Plasmid (m2) | sc-424640-NIC-2 | 20 µg | $410.00 |
Mouse Cisd1 encodes mitoNEET, a [2Fe–2S] cluster–binding protein anchored to the outer mitochondrial membrane that participates in iron–sulfur cluster transfer, redox regulation, and coordination of mitochondrial bioenergetics with cellular metabolic state. mitoNEET influences mitochondrial morphology and oxidative capacity, interfacing with pathways that govern reactive oxygen species handling, lipid utilization, and iron homeostasis. Dysregulation of CISD1/mitoNEET-associated processes has been linked in the literature to metabolic dysfunction, mitochondrial stress responses, and neurodegeneration-relevant phenotypes, making it a useful node for mechanistic studies of mitochondrial signaling. In mouse models, perturbation of Cisd1 can be leveraged to interrogate how mitochondrial outer membrane redox control shapes cellular physiology in tissues with high metabolic demand.
mitoNEET Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Cisd1 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Cisd1. 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 Cisd1 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 Cisd1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.