TLR5 CRISPR Activation Plasmid (h): sc-401154-ACT

Human TLR5 encodes Toll-like receptor 5, a pattern-recognition receptor that detects bacterial flagellin and initiates innate immune signaling at the plasma membrane. Upon ligand engagement, TLR5 signals primarily through the MYD88-dependent axis to activate NF-κB and MAPK pathways, promoting transcription of pro-inflammatory cytokines and chemokines that shape mucosal and systemic immune responses. TLR5 activity influences epithelial barrier function and host–microbiome interactions, and altered signaling has been linked to inflammatory phenotypes and susceptibility to infection. Because of its central role in pathogen sensing and immune tone, TLR5 is frequently studied in immunology, microbiology, and inflammation-focused cell models.

TLR5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TLR5 expression without altering the underlying DNA sequence.

TLR5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TLR5 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 TLR5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TLR5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TLR5 locus and enabling the study of TLR5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TLR5 pathway restoration in tumor cells with silenced or reduced TLR5 expression.

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