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.
Bad Double Nickase Plasmid (h): sc-400419-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Bad 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
- Bad Double Nickase Plasmid (h) and Bad Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting BAD. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Bad Antibody (C-7): sc-8044
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Bad Double Nickase Plasmid (h) | sc-400419-NIC | 20 µg | $410.00 | |||
Bad Double Nickase Plasmid (h2) | sc-400419-NIC-2 | 20 µg | $410.00 |
Human BAD encodes Bad, a BH3-only pro-apoptotic member of the BCL-2 family that integrates survival and death signals at the mitochondrial outer membrane. Bad promotes apoptosis by binding and neutralizing anti-apoptotic proteins such as BCL-2 and BCL-XL, thereby facilitating BAX/BAK-dependent mitochondrial outer membrane permeabilization and caspase activation. Its activity is tightly regulated by phosphorylation downstream of PI3K–AKT and MAPK signaling, which can sequester Bad via 14-3-3 proteins and shift cells toward survival. Dysregulated BAD signaling has been implicated in altered apoptotic thresholds relevant to tumor biology, neurodegeneration, and stress responses, making it a useful node for studying mitochondrial checkpoint control.
Bad Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the BAD locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within BAD. 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 BAD 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 BAD-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.