Rap1GAP Double Nickase Plasmid (h): sc-403721-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Rap1GAP 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
- Rap1GAP Double Nickase Plasmid (h) and Rap1GAP Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RAP1GAP. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Rap1GAP Antibody (D-9): sc-166586
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Rap1GAP Double Nickase Plasmid (h) | sc-403721-NIC | 20 µg | $410.00 | |||
Rap1GAP Double Nickase Plasmid (h2) | sc-403721-NIC-2 | 20 µg | $410.00 |
RAP1GAP encodes Rap1 GTPase-activating protein (Rap1GAP), a negative regulator of the small GTPase Rap1 that accelerates GTP hydrolysis to restrain Rap1 signaling. By modulating Rap1-dependent pathways controlling integrin activation, cell–cell junction dynamics, and actin cytoskeleton remodeling, Rap1GAP influences adhesion, migration, and polarized cell behavior. Altered RAP1GAP expression or activity has been reported across multiple disease contexts, where dysregulated Rap1 signaling can perturb MAPK/ERK and PI3K-linked networks and remodel extracellular matrix interactions. As a signaling brake at the interface of membrane receptors and downstream GTPase circuits, RAP1GAP is frequently studied to dissect mechanisms of invasion, barrier function, and metastatic-like phenotypes in model systems.
Rap1GAP Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RAP1GAP locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RAP1GAP. 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 RAP1GAP 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 RAP1GAP-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.