Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
C3G Double Nickase Plasmid (h): sc-401616-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- C3G 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
- C3G Double Nickase Plasmid (h) and C3G Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RAPGEF1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: C3G Antibody (G-4): sc-17840
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
C3G Double Nickase Plasmid (h) | sc-401616-NIC | 20 µg | $410.00 | |||
C3G Double Nickase Plasmid (h2) | sc-401616-NIC-2 | 20 µg | $410.00 |
RAPGEF1 encodes C3G, a guanine nucleotide exchange factor that activates Rap1 and related small GTPases to coordinate integrin-dependent adhesion, cytoskeletal remodeling, and cell migration. Through adaptor-mediated recruitment downstream of receptor tyrosine kinases and immune receptors, C3G helps couple extracellular cues to MAPK and Rap1 signaling that influence proliferation, differentiation, and vesicular trafficking. In hematopoietic and epithelial contexts, perturbation of RAPGEF1 has been linked to altered cell–cell junction dynamics and dysregulated signaling networks relevant to oncogenic transformation and immune cell function. As a pathway node connecting tyrosine phosphorylation to Rap-driven responses, C3G is frequently studied in models of invasion, leukocyte activation, and receptor-driven signaling plasticity.
C3G Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RAPGEF1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RAPGEF1. 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 RAPGEF1 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 RAPGEF1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.