RGNEF Double Nickase Plasmid (h): sc-404155-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RGNEF 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
- RGNEF Double Nickase Plasmid (h) and RGNEF Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ARHGEF28. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RGNEF Double Nickase Plasmid (h) | sc-404155-NIC | 20 µg | $410.00 | |||
RGNEF Double Nickase Plasmid (h2) | sc-404155-NIC-2 | 20 µg | $410.00 |
ARHGEF28 encodes RGNEF, a Rho guanine nucleotide exchange factor that activates Rho family GTPases to coordinate actin cytoskeleton remodeling, cell adhesion dynamics, and directional migration. Through coupling to RhoA-dependent signaling and downstream effectors such as ROCK and myosin contractility modules, RGNEF helps tune focal adhesion turnover and cytoskeletal tension. RGNEF also contains RNA-binding features that have been linked to stress granule biology and regulation of mRNA fate, connecting cytoskeletal signaling with post-transcriptional control. Dysregulation of ARHGEF28/RGNEF-associated pathways has been investigated in contexts including altered motility programs and neurodegeneration-relevant RNA/protein aggregation mechanisms, supporting its use as a research target in pathway and phenotype studies.
RGNEF Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ARHGEF28 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ARHGEF28. 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 ARHGEF28 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 ARHGEF28-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.