MARCH8 Double Nickase Plasmid (m): sc-428061-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MARCH8 Double Nickase Plasmid (m) 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
- MARCH8 Double Nickase Plasmid (m) and MARCH8 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting March8. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MARCH8 Double Nickase Plasmid (m) | sc-428061-NIC | 20 µg | $410.00 | |||
MARCH8 Double Nickase Plasmid (m2) | sc-428061-NIC-2 | 20 µg | $410.00 |
March8 encodes MARCH8, a membrane-associated RING-CH E3 ubiquitin ligase that localizes primarily to endosomal and trans-Golgi membranes and regulates the ubiquitination-dependent trafficking and turnover of multiple cell-surface and intracellular proteins. By promoting ubiquitin-mediated sorting to lysosomal degradation, MARCH8 helps control receptor abundance, immune signaling outputs, and membrane protein quality control within the endolysosomal system. In mouse models, MARCH8 has been linked to pathways governing antigen presentation, innate immune restriction of enveloped viruses, and modulation of signaling receptors through ubiquitin–proteasome and lysosome-associated processes. Dysregulated ubiquitination and membrane trafficking are broadly relevant to inflammatory phenotypes and pathogen–host interactions, making March8 a useful locus for mechanistic studies of immune regulation and protein homeostasis.
MARCH8 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the March8 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within March8. 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 March8 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 March8-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.