Dectin-1 CRISPR Activation Plasmid (h): sc-417053-ACT

Human CLEC7A encodes Dectin-1, a C-type lectin receptor prominently expressed by myeloid cells that recognizes β-glucans on fungal cell walls and selected endogenous ligands. Upon ligand engagement, Dectin-1 signals through an ITAM-like motif to activate SYK, CARD9–BCL10–MALT1, NF-κB, and MAPK pathways, coordinating phagocytosis, respiratory burst, cytokine production, and inflammasome-associated responses. This receptor also shapes cross-talk with Toll-like receptors and influences antigen presentation and Th17-skewing innate-adaptive communication. Altered CLEC7A/Dectin-1 activity has been linked to susceptibility to fungal infection, dysregulated inflammatory signaling, and tumor–myeloid microenvironment phenotypes relevant to immunology and host–pathogen research.

Dectin-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CLEC7A expression without altering the underlying DNA sequence.

Dectin-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CLEC7A 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 CLEC7A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Dectin-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CLEC7A locus and enabling the study of Dectin-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Dectin-1 pathway restoration in tumor cells with silenced or reduced CLEC7A expression.

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