HLA-DMα Double Nickase Plasmid (h): sc-404576-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HLA-DMα 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
- HLA-DMα Double Nickase Plasmid (h) and HLA-DMα Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting HLA-DMA. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HLA-DMα Antibody (NB-B33): sc-134356
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HLA-DMα Double Nickase Plasmid (h) | sc-404576-NIC | 20 µg | $410.00 | |||
HLA-DMα Double Nickase Plasmid (h2) | sc-404576-NIC-2 | 20 µg | $410.00 |
HLA-DMA encodes the HLA-DM alpha chain, a non-classical MHC class II molecule that forms a heterodimer with HLA-DMB in late endosomal/lysosomal compartments of antigen-presenting cells. HLA-DM catalyzes peptide exchange on MHC class II by removing CLIP from HLA-DR/DP/DQ and stabilizing high-affinity peptide loading, shaping the immunopeptidome presented to CD4+ T cells. This activity links HLA-DMA to antigen processing and presentation pathways, endosomal trafficking, and adaptive immune activation. Genetic variation or altered expression within the HLA class II region, including components of the HLA-DM system, is associated with immune dysregulation and susceptibility to autoimmune and inflammatory disease phenotypes.
HLA-DMα Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the HLA-DMA locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within HLA-DMA. 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 HLA-DMA 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 HLA-DMA-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.