CD1D CRISPR Activation Plasmid (h): sc-402435-ACT

CD1D encodes CD1d, a non-classical MHC class I–like antigen-presenting molecule that binds lipid and glycolipid antigens and displays them at the cell surface for recognition by invariant natural killer T (iNKT) cells. By shaping iNKT-cell activation and downstream cytokine programs, CD1d contributes to immune surveillance, tolerance, and inflammatory regulation across tissues. CD1d-dependent lipid antigen presentation intersects with endosomal trafficking, antigen processing, and innate-like lymphocyte signaling pathways that influence tumor immunology, infection biology, and chronic inflammatory states. Altered CD1D expression or function has been associated with dysregulated immune responses observed in autoimmunity, metabolic inflammation, and cancer microenvironments.

CD1D CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CD1D expression without altering the underlying DNA sequence.

CD1D CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CD1D 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 CD1D transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD1D expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CD1D locus and enabling the study of CD1D-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD1D pathway restoration in tumor cells with silenced or reduced CD1D expression.

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