CD1A CRISPR Activation Plasmid (h): sc-401734-ACT

CD1A encodes a non-classical MHC class I–like antigen-presenting molecule expressed prominently on dendritic cells, including Langerhans cells, where it binds and displays lipid and glycolipid antigens to T cells. By shaping lipid antigen presentation and T cell priming, CD1A contributes to immune surveillance at epithelial barriers and influences cytokine programs that modulate adaptive immune responses. Altered CD1A expression and CD1A-restricted T cell activity have been associated with inflammatory skin disorders and immune dysregulation, and CD1A is frequently used as a marker for dendritic cell differentiation and maturation states. In biomedical research, perturbing CD1A helps dissect lipid-driven immunological pathways and antigen-presenting cell function in human systems.

CD1A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CD1A expression without altering the underlying DNA sequence.

CD1A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CD1A 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 CD1A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD1A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CD1A locus and enabling the study of CD1A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD1A pathway restoration in tumor cells with silenced or reduced CD1A expression.

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