ARF6 Double Nickase Plasmid (h): sc-400799-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ARF6 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
- ARF6 Double Nickase Plasmid (h) and ARF6 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ARF6. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ARF6 Antibody (3A-1): sc-7971
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ARF6 Double Nickase Plasmid (h) | sc-400799-NIC | 20 µg | $410.00 | |||
ARF6 Double Nickase Plasmid (h2) | sc-400799-NIC-2 | 20 µg | $410.00 |
ADP-ribosylation factor 6 (ARF6) is a small GTPase that regulates membrane trafficking, actin cytoskeleton remodeling, and phosphoinositide signaling at the plasma membrane and endosomal compartments. By cycling between GDP- and GTP-bound states, ARF6 coordinates clathrin-independent endocytosis, recycling of membrane proteins, and integrin-dependent cell adhesion and migration, interfacing with pathways involving PI(4,5)P₂ dynamics and Rho family GTPases. ARF6-dependent vesicle transport influences receptor internalization and downstream signaling output, shaping processes such as cell polarity and cytokinesis. Dysregulated ARF6 activity has been linked to altered invasive behavior and metastatic signaling programs in cancer biology, and it is also implicated in host–pathogen entry mechanisms and neurodevelopmental processes relevant to disease models.
ARF6 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ARF6 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ARF6. 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 ARF6 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 ARF6-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.