TLR6 Knockout HEK293T Cell Line

The TLR6 Knockout HEK293T Cell Line is a CRISPR/Cas9-edited knockout cell line engineered for targeted disruption of the TLR6 gene in the HEK293T human embryonic kidney epithelial cell line. This stable knockout cell line provides a defined loss-of-function model for investigating TLR6-mediated innate immune signaling. By eliminating expression of full-length TLR6, the line enables precise dissection of TLR6-dependent pathways without interference from other closely related Toll-like receptors. Researchers can use this model for reconstitution studies, validating ligand specificity, and screening modulators of the TLR2/TLR6 heterodimer.

HEK293T cells are a widely adopted host for cell biology studies due to their highly transfectable nature and expression of SV40 large T-antigen, which supports episomal plasmid replication. Originating from human embryonic kidney epithelium, these cells offer a relevant epithelial context for innate immunity research. Their robust growth characteristics and extensive molecular characterization make them a reliable platform for signaling pathway analysis, protein interaction studies, and functional genomics applications requiring controlled genetic manipulation.

TLR6 encodes a pattern recognition receptor that heterodimerizes with TLR2 to detect diacylated lipopeptides from pathogens, including MALP-2, FSL-1, and Pam2CSK4. Ligand engagement recruits the adaptors TIRAP and MyD88, which in turn activate IRAK4 and TRAF6. This triggers downstream phosphorylation cascades through the IKK complex and MAP kinases, activating transcription factors NF-??B and AP-1. Consequently, TLR6 signaling drives expression of pro-inflammatory cytokines such as TNF-??, IL-6, and IL-8. Co-receptors CD36 and CD14 facilitate ligand presentation, enhancing the sensitivity of this pathogen-sensing pathway.

In the HEK293T background, TLR6 knockout offers a clean cellular system to study TLR6 function, as these cells express low levels of endogenous TLRs. The loss-of-function model is particularly useful for discriminating between signaling initiated by TLR2/TLR6 versus TLR2/TLR1 heterodimers. It enables robust analysis of TLR6-dependent protein?Cprotein interactions, signal transduction kinetics, and transcriptional responses. This knockout line also serves as a reliable control for TLR6-targeted pharmacological inhibitors and for experiments requiring complementation with wild-type or mutant TLR6 constructs.

Applications include NF-??B luciferase reporter assays, ELISA-based cytokine quantification, Western blotting for phospho-signaling intermediates, co-immunoprecipitation of receptor complexes, and RT-qPCR for downstream gene expression. The line is ideal for host?Cpathogen interaction studies, innate immune pathway dissection, and high-throughput screening of immunomodulatory compounds. For more information, please contact Ascent Research.