p35 Double Nickase Plasmid (h): sc-400431-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- p35 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
- p35 Double Nickase Plasmid (h) and p35 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CDK5R1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: p35 Antibody (B-1): sc-518009
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
p35 Double Nickase Plasmid (h) | sc-400431-NIC | 20 µg | $410.00 | |||
p35 Double Nickase Plasmid (h2) | sc-400431-NIC-2 | 20 µg | $410.00 |
CDK5R1 encodes p35, a neuron-enriched regulatory subunit that binds and activates cyclin-dependent kinase 5 (CDK5), directing kinase activity toward substrates that control cytoskeletal remodeling, neurite outgrowth, synaptic vesicle trafficking, and neuronal migration. Through CDK5/p35 signaling, CDK5R1 influences phosphorylation networks that intersect with MAPK signaling, axon guidance, and neurodevelopmental programs. Dysregulated p35 processing and altered CDK5 activity have been associated with neuronal stress responses and aberrant phosphorylation events implicated in neurodegenerative and neurodevelopmental disease mechanisms. Accordingly, CDK5R1 is widely studied for its role in maintaining neuronal structure and function under physiological and pathological conditions.
p35 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CDK5R1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CDK5R1. 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 CDK5R1 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 CDK5R1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.