TLR8 CRISPR Activation Plasmid (h): sc-401721-ACT

Human TLR8 (Toll-like receptor 8) is an endosomal pattern-recognition receptor that detects uridine-rich single-stranded RNA from pathogens and damaged cells, initiating innate immune signaling. Upon activation, TLR8 engages MyD88-dependent cascades that drive NF-κB and IRF-mediated transcriptional programs, promoting inflammatory cytokine and chemokine expression and shaping antigen-presenting cell function. TLR8 activity influences monocyte/macrophage and dendritic cell responses and intersects with interferon and inflammasome-related pathways that coordinate antimicrobial defense. Dysregulated TLR8 signaling has been linked to aberrant inflammatory states and immune-mediated disease mechanisms, making it a useful node for studying innate immune regulation and host–pathogen interactions.

TLR8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TLR8 expression without altering the underlying DNA sequence.

TLR8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TLR8 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 TLR8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TLR8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TLR8 locus and enabling the study of TLR8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TLR8 pathway restoration in tumor cells with silenced or reduced TLR8 expression.

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