MARCH5 Double Nickase Plasmid (h): sc-404655-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MARCH5 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
- MARCH5 Double Nickase Plasmid (h) and MARCH5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting MARCH5. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MARCH5 Double Nickase Plasmid (h) | sc-404655-NIC | 20 µg | $410.00 | |||
MARCH5 Double Nickase Plasmid (h2) | sc-404655-NIC-2 | 20 µg | $410.00 |
MARCH5 (also known as MITOL) is an outer mitochondrial membrane RING-type E3 ubiquitin ligase that regulates mitochondrial protein quality control and organelle dynamics. By ubiquitinating substrates involved in mitochondrial fusion–fission balance and stress signaling, MARCH5 influences processes including mitophagy, apoptosis, and the integrated cellular response to mitochondrial dysfunction. It interfaces with ubiquitin–proteasome pathways and mitochondrial homeostasis networks that shape redox balance and bioenergetic adaptation. Dysregulated MARCH5 activity or expression has been linked in the literature to altered mitochondrial morphology and stress susceptibility in contexts relevant to neurodegeneration and cancer cell metabolism.
MARCH5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the MARCH5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within MARCH5. 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 MARCH5 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 MARCH5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.