HLA-DP β1 CRISPR Activation Plasmid (h): sc-401413-ACT

HLA-DPB1 encodes the HLA-DP β1 chain, a core component of MHC class II heterodimers that present extracellularly derived peptides to CD4+ T cells and shape adaptive immune recognition. Expression is concentrated in professional antigen-presenting cells and is regulated by cytokine-driven programs, including interferon-γ–responsive transcription, supporting antigen processing and presentation pathways and T cell priming. Variation in HLA-DPB1 alleles influences peptide-binding repertoires and immune responsiveness, linking this locus to differential susceptibility and outcomes in autoimmune and inflammatory conditions, infectious disease responses, and transplant-related alloreactivity. As part of the HLA class II region, HLA-DP β1 also serves as a key marker for dissecting antigen presentation mechanisms and immunogenetic associations in human immune biology.

HLA-DP β1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HLA-DPB1 expression without altering the underlying DNA sequence.

HLA-DP β1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HLA-DPB1 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 HLA-DPB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HLA-DP β1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HLA-DPB1 locus and enabling the study of HLA-DP β1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HLA-DP β1 pathway restoration in tumor cells with silenced or reduced HLA-DPB1 expression.

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