PCNA Double Nickase Plasmid (h): sc-400037-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PCNA 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
- PCNA Double Nickase Plasmid (h) and PCNA Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PCNA. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PCNA Antibody (PC10): sc-56
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PCNA Double Nickase Plasmid (h) | sc-400037-NIC | 20 µg | $410.00 | |||
PCNA Double Nickase Plasmid (h2) | sc-400037-NIC-2 | 20 µg | $410.00 |
PCNA (proliferating cell nuclear antigen) is a conserved DNA sliding clamp that encircles duplex DNA to increase the processivity of replicative polymerases and coordinate assembly of replication and repair complexes. It functions as a central platform for proteins bearing PIP-box motifs and integrates signaling through post-translational modifications, including ubiquitination and SUMOylation, to direct translesion synthesis, template switching, and repair pathway choice. PCNA is essential for S-phase progression, Okazaki fragment maturation, and multiple DNA damage response processes, linking replication stress to checkpoint control and chromatin restoration. Dysregulated PCNA-dependent replication and repair programs are broadly associated with genome instability phenotypes studied across cancer, neurodegeneration, and other disorders of DNA maintenance.
PCNA Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PCNA locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PCNA. 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 PCNA 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 PCNA-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.