DC-SIGNR CRISPR Activation Plasmid (h): sc-404217-ACT

Human CLEC4M encodes DC-SIGNR, a C-type lectin receptor enriched on endothelial cells that recognizes high-mannose and fucosylated glycans to mediate Ca2+-dependent carbohydrate binding. Through ligand capture and adhesion functions, DC-SIGNR influences cell–cell interactions and pathogen engagement at mucosal and vascular interfaces, with downstream effects on endocytic trafficking and innate immune signaling. CLEC4M activity intersects with glycan-dependent recognition processes that shape antigen handling, inflammatory responses, and leukocyte tethering. Genetic and expression variability in CLEC4M has been examined in contexts involving infectious disease susceptibility and immune-related phenotypes, supporting its relevance as a modulator of host–pathogen and host–microbe interactions.

DC-SIGNR CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CLEC4M expression without altering the underlying DNA sequence.

DC-SIGNR CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CLEC4M 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 CLEC4M transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DC-SIGNR expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CLEC4M locus and enabling the study of DC-SIGNR-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DC-SIGNR pathway restoration in tumor cells with silenced or reduced CLEC4M expression.

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