Akt1 Double Nickase Plasmid (h): sc-400014-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Akt1 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
- Akt1 Double Nickase Plasmid (h) and Akt1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting AKT1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Akt1 Antibody (G-5): sc-55523
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Akt1 Double Nickase Plasmid (h) | sc-400014-NIC | 20 µg | $410.00 | |||
Akt1 Double Nickase Plasmid (h2) | sc-400014-NIC-2 | 20 µg | $410.00 |
AKT1 encodes the serine/threonine kinase Akt1, a central effector of PI3K signaling that integrates growth factor and insulin inputs to regulate cell survival, proliferation, metabolism, and protein synthesis. Akt1 phosphorylates diverse substrates controlling apoptosis, cell-cycle progression, and mTORC1-mediated translation, and it coordinates cellular responses to oxidative stress and nutrient availability. Dysregulated AKT1 signaling is linked to oncogenic transformation and therapy resistance, and altered Akt1 activity has also been implicated in metabolic and neurodevelopmental phenotypes. In human cells, AKT1 pathway perturbations are frequently studied in the context of receptor tyrosine kinase activation, PTEN loss, and downstream FOXO, GSK3, and TSC2 signaling nodes.
Akt1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the AKT1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within AKT1. 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 AKT1 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 AKT1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.