GPR54 Double Nickase Plasmid (h): sc-403766-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GPR54 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
- GPR54 Double Nickase Plasmid (h) and GPR54 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting KISS1R. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GPR54 Double Nickase Plasmid (h) | sc-403766-NIC | 20 µg | $410.00 | |||
GPR54 Double Nickase Plasmid (h2) | sc-403766-NIC-2 | 20 µg | $410.00 |
KISS1R encodes the G protein–coupled receptor GPR54 (also known as KISS1R), a high-affinity receptor for kisspeptins that regulates GnRH neuron activity and links metabolic and developmental cues to activation of the hypothalamic–pituitary–gonadal axis. Upon ligand engagement, GPR54 primarily signals through Gαq/11 to stimulate phospholipase C, intracellular Ca²⁺ mobilization, and downstream MAPK/ERK pathway activity, influencing gene expression and neuroendocrine secretion. Altered KISS1R signaling is associated with reproductive endocrine phenotypes including hypogonadotropic hypogonadism and disorders of pubertal timing, and it has been studied for roles in cell migration and invasion programs in diverse model systems. These features make KISS1R a useful target for investigating GPCR-driven calcium signaling, neuroendocrine regulation, and pathway crosstalk in human cells.
GPR54 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the KISS1R locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within KISS1R. 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 KISS1R 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 KISS1R-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.