Rho GAP p190-B Double Nickase Plasmid (h): sc-405755-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Rho GAP p190-B 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
- Rho GAP p190-B Double Nickase Plasmid (h) and Rho GAP p190-B Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ARHGAP5. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Rho GAP p190-B Antibody (G-11): sc-393241
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Rho GAP p190-B Double Nickase Plasmid (h) | sc-405755-NIC | 20 µg | $410.00 | |||
Rho GAP p190-B Double Nickase Plasmid (h2) | sc-405755-NIC-2 | 20 µg | $410.00 |
ARHGAP5 encodes Rho GTPase-activating protein p190-B, a key negative regulator of Rho-family GTPases that accelerates GTP hydrolysis to restrain RhoA-dependent signaling. Through modulation of actin cytoskeleton dynamics, focal adhesion turnover, and contractility, p190-B influences cell shape, adhesion, migration, and cytokinesis, integrating cues from integrins and growth factor pathways. ARHGAP5 activity impacts downstream effectors such as ROCK and myosin light chain phosphorylation, linking Rho signaling to mechanotransduction and cell motility programs. Dysregulated Rho GTPase signaling involving ARHGAP5 has been associated with altered invasive behavior and aberrant tissue remodeling in cancer and other pathologies characterized by cytoskeletal imbalance.
Rho GAP p190-B Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ARHGAP5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ARHGAP5. 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 ARHGAP5 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 ARHGAP5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.