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.
GRP 75 Double Nickase Plasmid (h): sc-400471-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GRP 75 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
- GRP 75 Double Nickase Plasmid (h) and GRP 75 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting HSPA9. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GRP 75 Antibody (D-9): sc-133137
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GRP 75 Double Nickase Plasmid (h) | sc-400471-NIC | 20 µg | $410.00 | |||
GRP 75 Double Nickase Plasmid (h2) | sc-400471-NIC-2 | 20 µg | $410.00 |
HSPA9 encodes GRP75 (mortalin), a mitochondrial HSP70 family chaperone that supports import, folding, and quality control of nuclear-encoded proteins within the mitochondrial matrix. GRP75 regulates mitochondrial proteostasis, membrane potential maintenance, and stress-adaptive responses, linking chaperone activity to oxidative phosphorylation efficiency and reactive oxygen species homeostasis. It also participates in mitochondrial–ER communication and calcium handling, integrating organelle crosstalk with cellular metabolism. Dysregulated HSPA9/GRP75 function has been associated with mitochondrial dysfunction phenotypes, altered stress tolerance, and cancer- and neurodegeneration-relevant pathways where proteostasis and bioenergetic remodeling are perturbed.
GRP 75 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the HSPA9 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within HSPA9. 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 HSPA9 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 HSPA9-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.