PIG-A Double Nickase Plasmid (m): sc-422226-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PIG-A Double Nickase Plasmid (m) 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
- PIG-A Double Nickase Plasmid (m) and PIG-A Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Piga. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PIG-A Antibody (H-6): sc-374194
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PIG-A Double Nickase Plasmid (m) | sc-422226-NIC | 20 µg | $410.00 | |||
PIG-A Double Nickase Plasmid (m2) | sc-422226-NIC-2 | 20 µg | $410.00 |
Mouse Piga encodes PIG-A, a catalytic subunit of the glycosylphosphatidylinositol (GPI)-N-acetylglucosaminyltransferase complex that initiates GPI-anchor biosynthesis in the endoplasmic reticulum. By enabling attachment of GPI anchors, PIG-A supports cell-surface presentation of diverse GPI-anchored proteins involved in membrane organization, signal transduction, cell–cell interactions, and immune-related processes. Disruption of PIG-A perturbs GPI-anchor assembly and can alter trafficking and stability of GPI-linked proteins, impacting pathways that depend on lipid-anchored receptors and enzymes. PIGA dysfunction is a central genetic lesion in paroxysmal nocturnal hemoglobinuria and is also implicated in congenital disorders of glycosylation, making Piga a useful locus for studying glycosylation-dependent phenotypes and selection-based genetics.
PIG-A Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Piga locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Piga. 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 Piga 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 Piga-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.