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.
HSPA5/BiP/GRP78 Double Nickase Plasmid (m): sc-420699-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HSPA5/BiP/GRP78 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
- HSPA5/BiP/GRP78 Double Nickase Plasmid (m) and HSPA5/BiP/GRP78 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Hspa5. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HSPA5/BiP/GRP78 Antibody (76-E6): sc-13539
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HSPA5/BiP/GRP78 Double Nickase Plasmid (m) | sc-420699-NIC | 20 µg | $410.00 |
Hspa5 encodes the ER-resident chaperone HSPA5/BiP/GRP78, a central regulator of protein folding quality control and proteostasis in mouse cells. HSPA5 binds nascent or misfolded polypeptides and coordinates the unfolded protein response (UPR) through interactions that modulate ER stress sensors such as PERK, IRE1, and ATF6. By influencing ER-associated degradation (ERAD), Ca²⁺ homeostasis, and secretory pathway throughput, it helps determine whether cells adapt to stress or undergo apoptosis. Dysregulated HSPA5 activity is frequently studied in contexts of metabolic stress, neurodegeneration, and oncogenic signaling where chronic ER stress and altered proteostasis contribute to disease-relevant phenotypes.
HSPA5/BiP/GRP78 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Hspa5 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Hspa5. 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 Hspa5 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 Hspa5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.