TLR6 CRISPR Activation Plasmid (h): sc-401996-ACT

Human TLR6 encodes a Toll-like receptor that functions as a pattern-recognition receptor in innate immunity, most prominently as a heterodimer with TLR2 to detect diacylated lipoproteins from bacteria and other microbial products. Ligand engagement activates MyD88-dependent signaling cascades that converge on NF-κB and MAPK pathways, promoting transcriptional programs that shape cytokine and chemokine production and coordinate early inflammatory responses. TLR6 activity influences antigen-presenting cell maturation and crosstalk between innate and adaptive immunity through modulation of costimulatory signals and interferon-associated gene networks. Dysregulated TLR6 signaling has been implicated in infection susceptibility and chronic inflammatory states, and it is frequently studied in the context of airway inflammation, metabolic inflammation, and microbiome-associated immune phenotypes.

TLR6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TLR6 expression without altering the underlying DNA sequence.

TLR6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TLR6 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 TLR6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TLR6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TLR6 locus and enabling the study of TLR6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TLR6 pathway restoration in tumor cells with silenced or reduced TLR6 expression.

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