Alix Double Nickase Plasmid (h): sc-400350-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Alix 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
- Alix Double Nickase Plasmid (h) and Alix Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PDCD6IP. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Alix Antibody (1A12): sc-53540
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Alix Double Nickase Plasmid (h) | sc-400350-NIC | 20 µg | $410.00 | |||
Alix Double Nickase Plasmid (h2) | sc-400350-NIC-2 | 20 µg | $410.00 |
PDCD6IP encodes Alix, an ESCRT-associated adaptor protein that coordinates membrane remodeling events including multivesicular body biogenesis, endosomal cargo sorting, and exosome biogenesis. Alix interacts with ESCRT-I/III components and partners such as TSG101 and CHMP4 to support cytokinetic abscission, plasma membrane repair, and receptor downregulation, linking it to growth factor and immune signaling outputs. Through its roles in endolysosomal trafficking and budding processes, Alix is frequently studied in the context of viral egress, autophagy-related membrane dynamics, and neuronal homeostasis. Dysregulation of PDCD6IP-dependent trafficking has been associated with altered extracellular vesicle signaling and proteostasis pathways relevant to cancer biology, neurodegeneration, and inflammatory states.
Alix Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PDCD6IP locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PDCD6IP. 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 PDCD6IP 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 PDCD6IP-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.