PKAα cat Double Nickase Plasmid (h): sc-400262-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PKAα cat 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
- PKAα cat Double Nickase Plasmid (h) and PKAα cat Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PRKACA. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PKAα cat Antibody (A-2): sc-28315
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PKAα cat Double Nickase Plasmid (h) | sc-400262-NIC | 20 µg | $410.00 | |||
PKAα cat Double Nickase Plasmid (h2) | sc-400262-NIC-2 | 20 µg | $410.00 |
PRKACA encodes the catalytic subunit alpha of cAMP-dependent protein kinase A (PKAα), a central effector of GPCR–adenylyl cyclase signaling that phosphorylates diverse substrates controlling metabolism, ion channel activity, transcription, and cell-cycle progression. PKAα regulates CREB-dependent gene expression and integrates with MAPK, calcium, and AKT pathway nodes to shape context-specific cellular responses. Aberrant PRKACA activity is linked to dysregulated cAMP signaling and has been implicated in endocrine and hepatic tumor biology, adrenal hyperfunction, and altered differentiation programs. As a core kinase within the cAMP/PKA axis, PRKACA is widely studied in signal transduction, stress responses, and phosphoproteomic network mapping.
PKAα cat Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PRKACA locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PRKACA. 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 PRKACA 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 PRKACA-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.