PI 3-kinase p85α Double Nickase Plasmid (h): sc-400224-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PI 3-kinase p85α 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
- PI 3-kinase p85α Double Nickase Plasmid (h) and PI 3-kinase p85α Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PIK3R1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PI 3-kinase p85α Antibody (B-9): sc-1637
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PI 3-kinase p85α Double Nickase Plasmid (h) | sc-400224-NIC | 20 µg | $410.00 | |||
PI 3-kinase p85α Double Nickase Plasmid (h2) | sc-400224-NIC-2 | 20 µg | $410.00 |
PIK3R1 encodes the p85α regulatory subunit of class IA phosphoinositide 3-kinase (PI3K), a key adaptor that stabilizes and recruits p110 catalytic subunits to activated receptor tyrosine kinases and IRS proteins. By coupling upstream growth factor signaling to PIP3 production, PI3K p85α controls AKT–mTOR and related pathways governing cell survival, proliferation, metabolism, and feedback regulation of insulin signaling. p85α also influences signaling complex assembly through SH2 domain–mediated phosphotyrosine binding and contributes to crosstalk with MAPK and other stress-response networks. Altered PIK3R1 function and pathway dysregulation are associated with oncogenic signaling states and metabolic phenotypes, making it a widely used node for mechanistic studies of PI3K pathway control.
PI 3-kinase p85α Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PIK3R1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PIK3R1. 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 PIK3R1 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 PIK3R1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.