Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Nir2 Double Nickase Plasmid (m): sc-422251-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Nir2 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
- Nir2 Double Nickase Plasmid (m) and Nir2 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Pitpnm1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Nir2 Antibody (8): sc-136140
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Nir2 Double Nickase Plasmid (m) | sc-422251-NIC | 20 µg | $410.00 | |||
Nir2 Double Nickase Plasmid (m2) | sc-422251-NIC-2 | 20 µg | $410.00 |
Pitpnm1 encodes Nir2, a phosphatidylinositol transfer protein that supports phosphoinositide homeostasis by exchanging lipids between membranes and sustaining phosphatidylinositol 4,5-bisphosphate pools at the plasma membrane. In mouse cells, Nir2 links receptor-driven phospholipase C signaling to lipid replenishment at endoplasmic reticulum–plasma membrane contact sites, influencing calcium-dependent signaling, membrane trafficking, and cytoskeletal dynamics. Through its role in regulating phosphoinositide turnover, Nir2 contributes to processes such as cell migration, neurite outgrowth, and vesicle transport that are sensitive to lipid signaling flux. Dysregulation of phosphoinositide metabolism and membrane contact site function is broadly relevant to studies of neurobiology, metabolism, and proliferative signaling, providing mechanistic entry points for disease-oriented research models.
Nir2 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Pitpnm1 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Pitpnm1. 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 Pitpnm1 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 Pitpnm1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.