ROS-GC1 Double Nickase Plasmid (h): sc-404483-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ROS-GC1 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
- ROS-GC1 Double Nickase Plasmid (h) and ROS-GC1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GUCY2D. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ROS-GC1 Antibody (B-7): sc-376217
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ROS-GC1 Double Nickase Plasmid (h) | sc-404483-NIC | 20 µg | $410.00 | |||
ROS-GC1 Double Nickase Plasmid (h2) | sc-404483-NIC-2 | 20 µg | $410.00 |
Human GUCY2D encodes retinal outer segment guanylate cyclase 1 (ROS-GC1), a membrane-associated enzyme that converts GTP to cGMP to sustain cyclic nucleotide signaling in photoreceptor cells. ROS-GC1 operates downstream of phototransduction, integrating Ca²⁺-dependent feedback via guanylate cyclase–activating proteins to restore cGMP levels and regulate cGMP-gated channel activity. This pathway supports outer segment homeostasis and synaptic signaling by coupling light-driven changes in intracellular Ca²⁺ to cyclic nucleotide turnover. Genetic perturbations in GUCY2D are strongly linked to inherited retinal degeneration phenotypes, making the gene a key node for studying cGMP metabolism, sensory signaling fidelity, and photoreceptor survival mechanisms.
ROS-GC1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GUCY2D locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GUCY2D. 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 GUCY2D 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 GUCY2D-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.