DNA-PKCS Double Nickase Plasmid (m): sc-422405-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DNA-PKCS Double Nickase Plasmid (m) 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
- DNA-PKCS Double Nickase Plasmid (m) and DNA-PKCS Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Prkdc. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: DNA-PKCS Antibody (G-12): sc-390849
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DNA-PKCS Double Nickase Plasmid (m) | sc-422405-NIC | 20 µg | $410.00 | |||
DNA-PKCS Double Nickase Plasmid (m2) | sc-422405-NIC-2 | 20 µg | $410.00 |
Prkdc encodes the catalytic subunit of DNA-dependent protein kinase, DNA-PKCS, a central regulator of DNA double-strand break repair through the non-homologous end joining (NHEJ) pathway. DNA-PKCS forms an active holoenzyme with Ku70/Ku80 to coordinate end recognition, synapsis, and end processing, and it participates in V(D)J recombination and class switch recombination in developing lymphocytes. As a core component of the DNA damage response, DNA-PKCS influences cell-cycle checkpoint signaling, genome stability, and cellular sensitivity to genotoxic stress. Altered PRKDC function is commonly used to model defects in NHEJ, chromosomal instability phenotypes, and mechanisms underlying immunodeficiency- and cancer-relevant DNA repair vulnerabilities in mouse systems.
DNA-PKCS Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Prkdc locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Prkdc. 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 Prkdc 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 Prkdc-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.