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.
Bak Double Nickase Plasmid (h): sc-400646-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Bak 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
- Bak Double Nickase Plasmid (h) and Bak Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting BAK1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Bak Antibody (AT38E2): sc-517390
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Bak Double Nickase Plasmid (h) | sc-400646-NIC | 20 µg | $410.00 | |||
Bak Double Nickase Plasmid (h2) | sc-400646-NIC-2 | 20 µg | $410.00 |
BAK1 encodes Bak, a pro-apoptotic BCL-2 family effector that localizes to the mitochondrial outer membrane and promotes mitochondrial outer membrane permeabilization following cellular stress. Upon activation, Bak undergoes conformational change and oligomerization, enabling cytochrome c release and downstream caspase activation within the intrinsic apoptosis pathway. Bak function is tightly regulated by interactions with BH3-only proteins and anti-apoptotic factors such as BCL-2 and BCL-XL, linking it to checkpoint control of cell survival and stress responses. Dysregulation of BAK1-mediated apoptosis has been implicated in altered cell death thresholds observed across cancer biology, neurodegeneration, and immune homeostasis research contexts.
Bak Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the BAK1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within BAK1. 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 BAK1 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 BAK1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.