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.
PUMA Double Nickase Plasmid (m): sc-431320-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PUMA 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
- PUMA Double Nickase Plasmid (m) and PUMA Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Bbc3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PUMAα Antibody (B-6): sc-377015
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PUMA Double Nickase Plasmid (m) | sc-431320-NIC | 20 µg | $410.00 | |||
PUMA Double Nickase Plasmid (m2) | sc-431320-NIC-2 | 20 µg | $410.00 |
Mouse Bbc3 encodes PUMA (p53 upregulated modulator of apoptosis), a BH3-only member of the BCL-2 family that promotes mitochondrial outer membrane permeabilization by neutralizing anti-apoptotic BCL-2 proteins and enabling BAX/BAK activation. PUMA is rapidly induced by cellular stress and integrates signals from p53-dependent DNA damage responses, growth factor withdrawal, and other apoptotic cues to engage intrinsic apoptosis and caspase activation. Through these pathways, Bbc3/PUMA helps regulate tissue homeostasis and elimination of damaged cells, and its dysregulation is frequently studied in contexts of tumor suppression, neurodegeneration, and immune cell survival. As a central node in apoptosis and stress-response networks, PUMA is commonly used to dissect mechanisms of cell fate decisions and checkpoint control in mouse models.
PUMA Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Bbc3 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Bbc3. 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 Bbc3 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 Bbc3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.