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.
CHIP/STUB1 Double Nickase Plasmid (h): sc-400815-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CHIP/STUB1 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
- CHIP/STUB1 Double Nickase Plasmid (h) and CHIP/STUB1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting STUB1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CHIP/STUB1 Antibody (G-2): sc-133066
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CHIP/STUB1 Double Nickase Plasmid (h) | sc-400815-NIC | 20 µg | $410.00 | |||
CHIP/STUB1 Double Nickase Plasmid (h2) | sc-400815-NIC-2 | 20 µg | $410.00 |
STUB1 encodes CHIP, a U-box E3 ubiquitin ligase and co-chaperone that couples HSP70/HSP90 client recognition to ubiquitination and proteasomal turnover, thereby regulating protein quality control. CHIP/STUB1 also interfaces with autophagy pathways and stress-response signaling to maintain proteostasis under conditions such as heat shock and oxidative stress. By controlling the stability of misfolded or damaged proteins, it influences cellular homeostasis, mitochondrial function, and apoptosis-related processes. Dysregulation or pathogenic variation in STUB1 is linked to neurodegeneration and ataxia phenotypes, making it a widely studied node in proteinopathy-relevant pathways.
CHIP/STUB1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the STUB1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within STUB1. 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 STUB1 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 STUB1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.