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.
P5CRL Double Nickase Plasmid (h): sc-406033-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- P5CRL 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
- P5CRL Double Nickase Plasmid (h) and P5CRL Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PYCRL. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: P5CRL Antibody (E-63): sc-100487
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
P5CRL Double Nickase Plasmid (h) | sc-406033-NIC | 20 µg | $410.00 |
Human PYCRL encodes pyrroline-5-carboxylate reductase–like protein (P5CRL), a cytosolic enzyme linked to proline biosynthesis and broader amino acid redox metabolism through NAD(P)H-dependent reduction of pyrroline-5-carboxylate. By influencing the proline–P5C cycle, PYCRL activity can affect cellular redox balance, mitochondrial–cytosolic metabolic coupling, and stress-adaptive processes that intersect with extracellular matrix regulation and proteostasis. Perturbation of proline metabolism is frequently connected to altered proliferation and oxidative stress responses, making PYCRL a relevant target for mechanistic studies of metabolic rewiring in disease-associated contexts. Investigating PYCRL also supports research into how amino acid metabolism integrates with central carbon pathways and cellular stress signaling.
P5CRL Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PYCRL locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PYCRL. 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 PYCRL 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 PYCRL-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.