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.
mEH Double Nickase Plasmid (h): sc-404239-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- mEH 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
- mEH Double Nickase Plasmid (h) and mEH Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EPHX1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: mEH Antibody (17): sc-135984
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
mEH Double Nickase Plasmid (h) | sc-404239-NIC | 20 µg | $410.00 | |||
mEH Double Nickase Plasmid (h2) | sc-404239-NIC-2 | 20 µg | $410.00 |
Human EPHX1 encodes microsomal epoxide hydrolase (mEH), an endoplasmic reticulum–associated enzyme that hydrolyzes reactive epoxides to less reactive diols and thereby shapes cellular handling of xenobiotics and endogenous lipid epoxides. mEH participates in phase I/II metabolic networks that intersect with oxidative stress responses, redox homeostasis, and inflammatory signaling, influencing downstream pathways linked to detoxification capacity. Variation in EPHX1 activity is studied in the context of differential susceptibility to chemical exposures and modulation of bioactive epoxide intermediates. In biomedical research, EPHX1 is frequently examined for its impact on metabolic phenotype, cellular stress tolerance, and pathway crosstalk affecting lipid mediator signaling.
mEH Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EPHX1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EPHX1. 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 EPHX1 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 EPHX1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.