PML Double Nickase Plasmid (h): sc-400145-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PML 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
- PML Double Nickase Plasmid (h) and PML Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PML. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PML Antibody (E-11): sc-377390
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PML Double Nickase Plasmid (h) | sc-400145-NIC | 20 µg | $410.00 | |||
PML Double Nickase Plasmid (h2) | sc-400145-NIC-2 | 20 µg | $410.00 |
Promyelocytic leukemia protein (PML) is a TRIM family scaffold that nucleates PML nuclear bodies, organizing proteins involved in transcriptional regulation, DNA damage responses, senescence, apoptosis, and intrinsic antiviral defense. PML is regulated by post-translational modification, including SUMOylation, which governs nuclear body assembly and recruitment of partners such as p53- and ATR/CHK-linked factors during genotoxic stress. Through these hubs, PML influences chromatin state, interferon-stimulated gene programs, and genome stability pathways that shape cellular fate decisions. Dysregulation of PML has been associated with altered stress signaling and malignant transformation contexts, including leukemia-relevant mechanisms involving nuclear body disruption.
PML Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PML locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PML. 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 PML 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 PML-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.