Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Mel-18 Double Nickase Plasmid (m): sc-423764-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Mel-18 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
- Mel-18 Double Nickase Plasmid (m) and Mel-18 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Pcgf2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Mel-18 Antibody (H-1): sc-515329
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Mel-18 Double Nickase Plasmid (m) | sc-423764-NIC | 20 µg | $410.00 | |||
Mel-18 Double Nickase Plasmid (m2) | sc-423764-NIC-2 | 20 µg | $410.00 |
Pcgf2 encodes Mel-18, a Polycomb group RING finger protein that functions within canonical PRC1 complexes to regulate chromatin compaction and transcriptional repression through ubiquitination of histone H2A and cooperation with PRC2-deposited H3K27me3. In mouse cells, Mel-18 contributes to stable gene silencing programs that shape developmental patterning, stem and progenitor cell state, and cell-cycle control by modulating accessibility at lineage-specifying loci. By influencing Polycomb-dependent networks, Pcgf2 impacts differentiation trajectories and epigenetic memory, processes frequently perturbed in models of oncogenic transformation and immune dysregulation. Genetic perturbation of Pcgf2 is therefore used to dissect PRC1 subunit specificity, chromatin topology changes, and context-dependent transcriptional outputs.
Mel-18 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Pcgf2 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Pcgf2. 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 Pcgf2 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 Pcgf2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.