R-Spondin3 Double Nickase Plasmid (h): sc-404472-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- R-Spondin3 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
- R-Spondin3 Double Nickase Plasmid (h) and R-Spondin3 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RSPO3. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
R-Spondin3 Double Nickase Plasmid (h) | sc-404472-NIC | 20 µg | $410.00 | |||
R-Spondin3 Double Nickase Plasmid (h2) | sc-404472-NIC-2 | 20 µg | $410.00 |
RSPO3 encodes R-Spondin3, a secreted agonist of canonical Wnt signaling that potentiates Frizzled/LRP receptor activity by binding LGR4/5/6 and modulating RNF43/ZNRF3-mediated turnover of Wnt receptors. Through this axis, R-Spondin3 regulates stem and progenitor cell maintenance, epithelial renewal, angiogenic programs, and tissue morphogenesis. RSPO3-dependent signaling influences β-catenin–driven transcriptional networks that shape cell fate decisions and proliferative responses. Dysregulated RSPO3 expression or pathway engagement has been implicated in aberrant Wnt pathway activity observed across multiple disease-relevant contexts, supporting its use as a mechanistic node in pathway and developmental biology studies.
R-Spondin3 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RSPO3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RSPO3. 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 RSPO3 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 RSPO3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.