DNA pol ε A Double Nickase Plasmid (m): sc-422339-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DNA pol ε A 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 pol ε A Double Nickase Plasmid (m) and DNA pol ε A Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Pole. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: DNA pol ε A Antibody (3C5.1): sc-12728
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DNA pol ε A Double Nickase Plasmid (m) | sc-422339-NIC | 20 µg | $410.00 |
Mouse Pole encodes the catalytic subunit A of DNA polymerase epsilon, a high-fidelity replicative polymerase essential for leading-strand DNA synthesis during S phase. DNA pol ε A coordinates with the CMG helicase and replication factor complexes to support replisome progression, Okazaki fragment processing coordination, and accurate genome duplication. Beyond replication, Pole contributes to DNA damage tolerance and replication stress responses, interfacing with checkpoint signaling pathways that preserve fork stability. Disruption of replicative polymerase function is tightly linked to genome instability phenotypes that are widely used to model mutational processes and cancer-relevant DNA replication defects in experimental systems.
DNA pol ε A Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Pole locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Pole. 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 Pole 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 Pole-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.