AMPK alpha 1 Double Nickase Plasmid (h): sc-400104-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AMPK alpha 1 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
- AMPK alpha 1 Double Nickase Plasmid (h) and AMPK alpha 1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PRKAA1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: AMPK alpha 1 Antibody (H-4): sc-398861
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AMPK alpha 1 Double Nickase Plasmid (h) | sc-400104-NIC | 20 µg | $410.00 | |||
AMPK alpha 1 Double Nickase Plasmid (h2) | sc-400104-NIC-2 | 20 µg | $410.00 |
PRKAA1 encodes the catalytic α1 subunit of AMP-activated protein kinase (AMPK), a central sensor of cellular energy status that is activated by rising AMP/ADP and upstream kinases such as LKB1 and CAMKK2. AMPKα1 coordinates metabolic adaptation by regulating glucose uptake and glycolysis, inhibiting anabolic programs through mTORC1 suppression, and promoting autophagy and mitochondrial homeostasis via phosphorylation of downstream effectors. Through these activities, AMPK signaling shapes responses to nutrient stress, hypoxia, and oxidative stress, influencing cell growth, lipid metabolism, and inflammatory signaling. Dysregulated PRKAA1/AMPKα1 activity has been associated with metabolic disease phenotypes and is frequently interrogated in cancer biology where metabolic reprogramming impacts proliferation and survival.
AMPK alpha 1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PRKAA1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PRKAA1. 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 PRKAA1 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 PRKAA1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.