GPI-PLD Double Nickase Plasmid (h): sc-404938-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GPI-PLD 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
- GPI-PLD Double Nickase Plasmid (h) and GPI-PLD Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GPLD1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GPI-PLD Antibody (D-10): sc-365096
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GPI-PLD Double Nickase Plasmid (h) | sc-404938-NIC | 20 µg | $410.00 | |||
GPI-PLD Double Nickase Plasmid (h2) | sc-404938-NIC-2 | 20 µg | $410.00 |
GPLD1 encodes glycosylphosphatidylinositol-specific phospholipase D1 (GPI-PLD), a secreted enzyme that hydrolyzes GPI anchors and regulates the release of GPI-anchored proteins from cell membranes into extracellular compartments. By modulating the membrane association of diverse GPI-anchored proteins, GPI-PLD can influence cell–cell signaling, immune-related processes, and lipid-linked protein trafficking across tissues. GPLD1 expression and circulating GPI-PLD activity have been examined in the context of metabolic and inflammatory biology, including links to insulin resistance, liver-associated pathways, and systemic immune modulation. These functional connections make GPLD1 a useful target for mechanistic studies of GPI-anchor turnover, extracellular enzyme activity, and proteome remodeling.
GPI-PLD Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GPLD1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GPLD1. 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 GPLD1 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 GPLD1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.