MECL-1 Double Nickase Plasmid (h2): sc-402101-NIC-2
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MECL-1 Double Nickase Plasmid (h2) 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
- MECL-1 Double Nickase Plasmid (h2) and MECL-1 Double Nickase Plasmid (h22) encode distinct paired gRNA designs targeting PSMB10. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: MECL-1 Antibody (C-2): sc-133236
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MECL-1 Double Nickase Plasmid (h2) | sc-402101-NIC-2 | 20 µg | $410.00 |
Human PSMB10 encodes the proteasome subunit beta type-10 (MECL-1), an inducible catalytic component of the immunoproteasome that replaces constitutive β subunits to reshape proteolytic cleavage preferences during inflammatory signaling. MECL-1 supports ubiquitin–proteasome–mediated protein turnover and enhances MHC class I antigen processing, linking interferon-driven responses to adaptive immune surveillance and modulation of NF-κB–associated proteostasis programs. Dysregulation of immunoproteasome composition or activity is implicated in altered antigen presentation and chronic immune activation observed in autoimmunity, infection, and tumor immune evasion contexts. Gene editing of PSMB10 enables mechanistic interrogation of immunoproteasome-dependent peptide repertoires, proteome quality control under cytokine stress, and immune pathway rewiring in human cell models.
MECL-1 Double Nickase Plasmid (h2) consists of a matched pair of plasmids engineered for high-specificity editing of the PSMB10 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PSMB10. 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 PSMB10 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 PSMB10-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.