Twinkle Double Nickase Plasmid (m): sc-432601-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Twinkle 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
- Twinkle Double Nickase Plasmid (m) and Twinkle Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Twnk. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Twinkle Double Nickase Plasmid (m) | sc-432601-NIC | 20 µg | $410.00 |
Mouse Twnk encodes Twinkle, a mitochondrial DNA helicase essential for mtDNA replication and maintenance within nucleoids. Twinkle functions with the mitochondrial replisome to support oxidative phosphorylation by preserving mitochondrial genome copy number and integrity, thereby influencing cellular energy metabolism and reactive oxygen species homeostasis. Perturbation of TWNK activity is linked to mitochondrial DNA depletion and multiple mtDNA deletion phenotypes, making this pathway central to studies of mitochondrial dysfunction in neuromuscular and neurodegenerative disease biology. Twnk is therefore a widely used node for investigating mitochondrial genome stability, replication stress responses, and downstream impacts on respiratory chain biogenesis.
Twinkle Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Twnk locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Twnk. 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 Twnk 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 Twnk-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.