P5CR Double Nickase Plasmid (h): sc-410971-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- P5CR 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
- P5CR Double Nickase Plasmid (h) and P5CR Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PYCR1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: P5CR Antibody (A-12): sc-518219
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
P5CR Double Nickase Plasmid (h) | sc-410971-NIC | 20 µg | $410.00 | |||
P5CR Double Nickase Plasmid (h2) | sc-410971-NIC-2 | 20 µg | $410.00 |
PYCR1 encodes pyrroline-5-carboxylate reductase 1 (P5CR), a mitochondrial enzyme that catalyzes the NAD(P)H-dependent conversion of pyrroline-5-carboxylate to proline, linking glutamate/ornithine metabolism to proline biosynthesis. Through its role in proline cycling, P5CR contributes to cellular redox balance, mitochondrial homeostasis, and metabolic adaptation under oxidative or nutrient stress. PYCR1 activity intersects with amino acid metabolism, reactive oxygen species buffering, and extracellular matrix-related processes that depend on proline availability. Genetic perturbation of PYCR1 has been associated with connective tissue and neurodevelopmental phenotypes, making it a useful node for studying metabolic regulation and stress-response pathways.
P5CR Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PYCR1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PYCR1. 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 PYCR1 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 PYCR1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.