HLA-DRβ5 Double Nickase Plasmid (h): sc-402985-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HLA-DRβ5 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-DRβ5 Double Nickase Plasmid (h) and HLA-DRβ5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting HLA-DRB5. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HLA-DRβ5 Antibody (4D12): sc-53889
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HLA-DRβ5 Double Nickase Plasmid (h) | sc-402985-NIC | 20 µg | $410.00 | |||
HLA-DRβ5 Double Nickase Plasmid (h2) | sc-402985-NIC-2 | 20 µg | $410.00 |
HLA-DRB5 encodes the HLA-DR β5 chain, a polymorphic component of MHC class II heterodimers that present extracellularly derived peptides to CD4+ T cells. By assembling with HLA-DRA in antigen-presenting cells, HLA-DRβ5 contributes to endosomal peptide loading, immune synapse formation, and downstream T cell activation programs that shape adaptive immune responses. Variation in MHC class II peptide-binding grooves influences antigen repertoire and immune tolerance, linking HLA-DR loci to susceptibility and stratification across multiple immune-mediated and inflammatory disease contexts. HLA-DRB5 is therefore widely studied in antigen presentation biology, HLA haplotype mapping, and mechanisms regulating APC–T cell communication.
HLA-DRβ5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the HLA-DRB5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within HLA-DRB5. 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-DRB5 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-DRB5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.