Parkin Double Nickase Plasmid (h): sc-400226-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Parkin 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
- Parkin Double Nickase Plasmid (h) and Parkin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PARK2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Parkin Antibody (PRK8): sc-32282
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Parkin Double Nickase Plasmid (h) | sc-400226-NIC | 20 µg | $410.00 | |||
Parkin Double Nickase Plasmid (h2) | sc-400226-NIC-2 | 20 µg | $410.00 |
PARK2 encodes Parkin, an RBR-family E3 ubiquitin ligase that coordinates ubiquitin-dependent protein turnover and mitochondrial quality control. In response to mitochondrial depolarization, Parkin is recruited downstream of PINK1 to ubiquitinate outer mitochondrial membrane substrates, promoting mitophagy and remodeling of mitochondrial dynamics. Through its roles in proteostasis, oxidative stress responses, and synaptic maintenance, dysregulated Parkin activity is linked to neuronal vulnerability and is frequently studied in the context of early-onset Parkinson’s disease and related neurodegenerative mechanisms. Parkin also intersects with inflammatory signaling and metabolic adaptation pathways, making it relevant for investigations of mitochondrial dysfunction across cell types.
Parkin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PARK2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PARK2. 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 PARK2 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 PARK2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.