RIP/Rab Double Nickase Plasmid (h): sc-405836-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RIP/Rab 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
- RIP/Rab Double Nickase Plasmid (h) and RIP/Rab Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting AGFG1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: RIP/Rab Antibody (H-2): sc-166651
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RIP/Rab Double Nickase Plasmid (h) | sc-405836-NIC | 20 µg | $410.00 |
AGFG1 (RIP/Rab) encodes an ArfGAP and FG-repeat–containing protein implicated in clathrin-mediated endocytosis and membrane trafficking, linking small GTPase signaling to cargo internalization and endosomal sorting. Through interactions with components of the endocytic machinery, RIP/Rab contributes to vesicle budding, receptor recycling, and spatial organization of signaling complexes at the plasma membrane. AGFG1-dependent trafficking can influence downstream pathways by regulating the abundance and localization of surface receptors and transporters. Dysregulation of endocytic control and Rab/Arf-mediated trafficking has been associated with altered cell signaling and cellular homeostasis, making AGFG1 a useful target for mechanistic studies in disease-relevant models.
RIP/Rab Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the AGFG1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within AGFG1. 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 AGFG1 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 AGFG1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.