CARD 9 CRISPR Activation Plasmid (h): sc-402214-ACT

Caspase recruitment domain-containing protein 9 (CARD9) is an adaptor protein predominantly expressed in myeloid lineages that couples C-type lectin receptor signaling to downstream innate immune responses. Upon sensing fungal and other microbial ligands, CARD9 integrates signals from receptors such as Dectin-1 and Dectin-2 to promote NF-κB and MAPK pathway activation, inflammasome-associated cytokine production, and shaping of Th17-skewing inflammatory programs. Genetic and functional perturbation of CARD9 has been linked to altered host–microbe interactions and dysregulated inflammation, with associations spanning susceptibility to invasive fungal infection and immune-mediated inflammatory phenotypes. As a signaling hub at the interface of pattern recognition and transcriptional control, CARD9 is widely studied in pathways governing myeloid activation, cytokine networks, and barrier immunity.

CARD 9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CARD9 expression without altering the underlying DNA sequence.

CARD 9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CARD9 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 CARD9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CARD 9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CARD9 locus and enabling the study of CARD 9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CARD 9 pathway restoration in tumor cells with silenced or reduced CARD9 expression.

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