Rab 5A Double Nickase Plasmid (h): sc-417949-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Rab 5A 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
- Rab 5A Double Nickase Plasmid (h) and Rab 5A Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RAB5A. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Rab 5A Antibody (C-3): sc-515401
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Rab 5A Double Nickase Plasmid (h) | sc-417949-NIC | 20 µg | $410.00 |
RAB5A encodes Rab 5A, a small GTPase that functions as a master regulator of early endocytosis and endosomal membrane trafficking. In its GTP-bound state, Rab 5A coordinates clathrin-mediated internalization, early endosome fusion, and cargo sorting by recruiting effectors that control vesicle tethering and phosphoinositide signaling. Through these roles it influences receptor turnover and signal transduction dynamics across pathways such as growth factor receptor trafficking and endosome maturation. Dysregulated RAB5A activity or expression has been linked to altered intracellular transport programs observed in cancer cell invasion, neurodegenerative phenotypes, and pathogen entry mechanisms, making it a useful target for mechanistic studies of membrane traffic and signaling.
Rab 5A Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RAB5A locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RAB5A. 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 RAB5A 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 RAB5A-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.