HSP70-1 Double Nickase Plasmid (h): sc-418088-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HSP70-1 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
- HSP70-1 Double Nickase Plasmid (h) and HSP70-1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting HSPA1A. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HSP70 Antibody (F-3): sc-373867
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HSP70-1 Double Nickase Plasmid (h) | sc-418088-NIC | 20 µg | $410.00 | |||
HSP70-1 Double Nickase Plasmid (h2) | sc-418088-NIC-2 | 20 µg | $410.00 |
HSPA1A encodes the human heat shock protein HSP70-1, an ATP-dependent molecular chaperone that promotes proteostasis by assisting nascent polypeptide folding, preventing aggregation, and facilitating refolding or degradation of stress-damaged proteins. HSP70-1 operates within the cellular heat shock response in coordination with HSF1 and co-chaperones such as DNAJ/HSP40 and BAG family proteins, influencing ubiquitin–proteasome and autophagy-linked quality control. By modulating protein homeostasis, antigen presentation, and stress signaling, HSP70-1 is broadly relevant to conditions characterized by proteotoxic stress, including neurodegeneration, cancer cell survival under stress, and inflammatory states.
HSP70-1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the HSPA1A locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within HSPA1A. 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 HSPA1A 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 HSPA1A-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.