RhoA Double Nickase Plasmid (h): sc-400052-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RhoA 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
- RhoA Double Nickase Plasmid (h) and RhoA Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RHOA. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: RhoA Antibody (26C4): sc-418
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RhoA Double Nickase Plasmid (h) | sc-400052-NIC | 20 µg | $410.00 | |||
RhoA Double Nickase Plasmid (h2) | sc-400052-NIC-2 | 20 µg | $410.00 |
RHOA encodes the small GTPase RhoA, a central regulator of actin cytoskeleton dynamics, cell shape, and contractility through cycling between GDP- and GTP-bound states. Active RhoA signals prominently via ROCK and mDia effectors to control stress fiber formation, focal adhesion turnover, cytokinesis, and mechanotransduction, with broader integration into GPCR and integrin-dependent pathways. RhoA activity influences cell migration, epithelial barrier function, and transcriptional responses linked to cytoskeletal tension. Dysregulated RHOA signaling and recurrent mutations have been reported across multiple cancer types and are also implicated in cardiovascular remodeling and neurodevelopmental processes, supporting its relevance in disease-associated signaling networks.
RhoA Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RHOA locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RHOA. 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 RHOA 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 RHOA-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.