ME2 Double Nickase Plasmid (h): sc-402900-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ME2 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
- ME2 Double Nickase Plasmid (h) and ME2 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ME2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ME2 Antibody (F-5): sc-514850
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ME2 Double Nickase Plasmid (h) | sc-402900-NIC | 20 µg | $410.00 | |||
ME2 Double Nickase Plasmid (h2) | sc-402900-NIC-2 | 20 µg | $410.00 |
Human ME2 encodes the mitochondrial NAD(P)-dependent malic enzyme (ME2), which catalyzes oxidative decarboxylation of malate to pyruvate with concomitant production of NADH or NADPH. By linking the tricarboxylic acid cycle to anaplerosis and redox homeostasis, ME2 influences mitochondrial metabolism, reactive oxygen species handling, and biosynthetic flux through pyruvate- and citrate-dependent pathways. Altered ME2 activity has been associated with metabolic reprogramming observed in proliferative states and is studied in the context of mitochondrial dysfunction, oxidative stress, and oncogenic metabolism.
ME2 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ME2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ME2. 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 ME2 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 ME2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.