CLEC-2D CRISPR Activation Plasmid (h): sc-412035-ACT

CLEC2D encodes CLEC-2D, a C-type lectin–like ligand displayed on the cell surface that modulates immune recognition through interactions with NK cell receptors, particularly NKG2D and CD161 (KLRB1). By shaping activating and inhibitory signaling at the immune synapse, CLEC-2D influences NK cell cytotoxicity, cytokine release, and broader immune surveillance within inflammatory and tumor-associated microenvironments. Its expression is dynamically regulated by cellular stress and differentiation states, linking CLEC2D to pathways controlling immune evasion, leukocyte crosstalk, and tissue homeostasis. Altered CLEC2D levels have been reported across multiple malignancies and immune-mediated conditions, supporting its use as a molecular handle to study immune escape, antigen-independent cytotoxicity, and immunoregulatory phenotypes.

CLEC-2D CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CLEC2D expression without altering the underlying DNA sequence.

CLEC-2D CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CLEC2D 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 CLEC2D transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CLEC-2D expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CLEC2D locus and enabling the study of CLEC-2D-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CLEC-2D pathway restoration in tumor cells with silenced or reduced CLEC2D expression.

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