PDZ-RhoGEF Double Nickase Plasmid (h): sc-403444-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PDZ-RhoGEF 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
- PDZ-RhoGEF Double Nickase Plasmid (h) and PDZ-RhoGEF Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ARHGEF11. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PDZ-RhoGEF Antibody (D-9): sc-166740
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PDZ-RhoGEF Double Nickase Plasmid (h) | sc-403444-NIC | 20 µg | $410.00 | |||
PDZ-RhoGEF Double Nickase Plasmid (h2) | sc-403444-NIC-2 | 20 µg | $410.00 |
ARHGEF11 encodes PDZ-RhoGEF, a RhoA-specific guanine nucleotide exchange factor that links activated Gα12/13-coupled GPCR signaling to Rho GTPase activation and downstream actomyosin contractility. Through regulation of cytoskeletal remodeling, stress fiber formation, focal adhesion dynamics, and cell polarity, PDZ-RhoGEF influences processes such as migration, adhesion, and epithelial barrier function. PDZ domain–mediated scaffolding and signaling crosstalk integrate inputs from receptor signaling and junctional complexes to tune Rho/ROCK pathway output. Dysregulated RhoA signaling and ARHGEF11-associated network alterations have been implicated in pathological cell motility and invasion phenotypes and broader disease-relevant signaling changes in cancer and inflammatory contexts.
PDZ-RhoGEF Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ARHGEF11 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ARHGEF11. 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 ARHGEF11 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 ARHGEF11-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.