HLA-E Double Nickase Plasmid (h): sc-403124-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HLA-E 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-E Double Nickase Plasmid (h) and HLA-E Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting HLA-E. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HLA-E Antibody (MEM-E/02): sc-51621
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HLA-E Double Nickase Plasmid (h) | sc-403124-NIC | 20 µg | $410.00 | |||
HLA-E Double Nickase Plasmid (h2) | sc-403124-NIC-2 | 20 µg | $410.00 |
HLA-E encodes a non-classical MHC class I molecule that presents a restricted peptide repertoire, including leader peptides from other HLA class I proteins, to regulate immune surveillance. Surface HLA-E interacts with inhibitory and activating receptors such as CD94/NKG2A and CD94/NKG2C on NK cells and subsets of T cells, shaping cytotoxic responses and tolerance. Through antigen presentation and immune checkpoint-like signaling, HLA-E contributes to control of inflammation, viral immune evasion strategies, and modulation of tumor immunogenicity in the tumor microenvironment. Altered HLA-E expression has been associated with immune escape phenotypes and disease contexts where NK/T-cell recognition is perturbed, making it a useful target for mechanistic immunology research.
HLA-E Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the HLA-E locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within HLA-E. 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-E 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-E-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.