GARNL4 Double Nickase Plasmid (h): sc-411668-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GARNL4 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
- GARNL4 Double Nickase Plasmid (h) and GARNL4 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RAP1GAP2. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GARNL4 Double Nickase Plasmid (h) | sc-411668-NIC | 20 µg | $410.00 |
RAP1GAP2 encodes GARNL4, a Rap GTPase-activating protein that accelerates GTP hydrolysis on Rap1/Rap2 to constrain small GTPase signaling at the plasma membrane and cytoskeleton. By modulating Rap-dependent control of integrin activation, cell adhesion, and actin remodeling, GARNL4 contributes to regulation of platelet and leukocyte signaling, trafficking, and stimulus-coupled responses. This RAP1 pathway intersects with MAPK/ERK and PI3K-linked networks through upstream receptors and adaptor complexes, shaping cellular activation thresholds and secretion dynamics. Dysregulated Rap signaling and altered GAP activity are frequently studied in contexts of abnormal hemostasis, inflammatory signaling, and cancer-associated changes in adhesion and migration.
GARNL4 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RAP1GAP2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RAP1GAP2. 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 RAP1GAP2 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 RAP1GAP2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.