HLA-E CRISPR Activation Plasmid (m): sc-437133-ACT

H2-T-ps encodes a mouse MHC class Ib molecule functionally analogous to HLA-E, specialized in presenting a restricted peptide repertoire to regulate innate and adaptive immune surveillance. By engaging CD94/NKG2 receptors on NK cells and subsets of T cells, HLA-E–like molecules help tune cytotoxic activation thresholds, linking antigen processing in the ER with immune checkpoint–like signaling. This axis integrates with interferon-driven pathways that modulate antigen presentation and cellular stress responses, influencing tissue homeostasis during inflammation. Dysregulated expression or peptide loading of nonclassical MHC I can alter NK cell education and tumor–immune interactions, making this gene relevant to studies of infection, autoimmunity, and cancer immunobiology in mouse models.

HLA-E CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous H2-T-ps expression without altering the underlying DNA sequence.

HLA-E CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the H2-T-ps locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the H2-T-ps transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HLA-E expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native H2-T-ps locus and enabling the study of HLA-E-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HLA-E pathway restoration in tumor cells with silenced or reduced H2-T-ps expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.