GCH-I Double Nickase Plasmid (h): sc-402386-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GCH-I 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
- GCH-I Double Nickase Plasmid (h) and GCH-I Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GCH1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GCH-I Antibody (G-8): sc-376483
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GCH-I Double Nickase Plasmid (h) | sc-402386-NIC | 20 µg | $410.00 | |||
GCH-I Double Nickase Plasmid (h2) | sc-402386-NIC-2 | 20 µg | $410.00 |
GCH1 encodes GTP cyclohydrolase I (GCH-I), the rate-limiting enzyme in de novo tetrahydrobiopterin (BH4) biosynthesis from GTP. BH4 is an essential redox-active cofactor for aromatic amino acid hydroxylases and all nitric oxide synthase isoforms, thereby linking GCH-I activity to catecholamine and serotonin production as well as nitric oxide signaling and cellular redox balance. Altered GCH1 function can perturb neurotransmitter metabolism and endothelial nitric oxide pathways, making it relevant to studies of neurometabolic homeostasis, vascular biology, and oxidative stress. Variants in GCH1 are associated with inherited neurotransmitter disorders and have been investigated in the context of pain sensitivity and neuropsychiatric phenotypes, supporting mechanistic research into genotype–pathway relationships.
GCH-I Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GCH1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GCH1. 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 GCH1 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 GCH1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.