AKAP 95 Double Nickase Plasmid (h): sc-403651-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AKAP 95 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
- AKAP 95 Double Nickase Plasmid (h) and AKAP 95 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting AKAP8. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: AKAP 95 Antibody (F-11): sc-390335
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AKAP 95 Double Nickase Plasmid (h) | sc-403651-NIC | 20 µg | $410.00 | |||
AKAP 95 Double Nickase Plasmid (h2) | sc-403651-NIC-2 | 20 µg | $410.00 |
AKAP8 encodes AKAP 95, a nuclear A-kinase anchoring protein that scaffolds PKA and additional regulatory factors within chromatin-associated compartments to coordinate phosphorylation-dependent control of gene expression. AKAP 95 participates in chromatin organization, RNA processing and splicing, and mitotic events such as chromosome condensation and cell-cycle progression, linking signaling to nuclear architecture. Through these roles, AKAP8 is frequently studied in pathways governing proliferation, DNA/RNA homeostasis, and stress-responsive transcriptional programs. Dysregulation of AKAP 95-associated nuclear signaling and splicing networks has been implicated in disease-relevant phenotypes including altered growth control and aberrant transcript processing in cancer biology.
AKAP 95 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the AKAP8 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within AKAP8. 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 AKAP8 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 AKAP8-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.