Rubicon Double Nickase Plasmid (h): sc-404222-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Rubicon 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
- Rubicon Double Nickase Plasmid (h) and Rubicon Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RUBCN. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Rubicon Double Nickase Plasmid (h) | sc-404222-NIC | 20 µg | $410.00 | |||
Rubicon Double Nickase Plasmid (h2) | sc-404222-NIC-2 | 20 µg | $410.00 |
RUBCN encodes Rubicon, a RUN domain–containing protein that acts as a key negative regulator of autophagy by modulating VPS34/PI3KC3 activity and autophagosome maturation. Rubicon also participates in endolysosomal trafficking and intersects with innate immune signaling through regulation of LC3-associated phagocytosis and related vesicular pathways. By controlling autophagic flux and lysosome-dependent clearance, Rubicon influences cellular homeostasis under stress conditions and shapes inflammatory outputs. Dysregulated RUBCN/Rubicon activity has been implicated in contexts where autophagy and phagosome processing are altered, including neurodegeneration, infection biology, metabolic dysfunction, and cancer-related cellular adaptation, making it a useful target for mechanistic studies.
Rubicon Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RUBCN locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RUBCN. 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 RUBCN 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 RUBCN-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.