NF-κB–SEAP reporter HEK293 cells expressing human TLR9
INVIVOGEN ALSO OFFERS:
• HEK-Blue mTLR9 cells
• HEK-Dual hTLR9 cells
• TLR9 ligands - Including stimulatory and inhibitory ODNs, as well as bacterial DNAInvivoGen offers a human embryonic kidney 293 (HEK293)-derived cell line, specifically designed for the study of human TLR9 (Toll-Like Receptor 9)-induced NF-κB signaling pathway:• HEK-Blue hTLR9 cellsHEK-Blue hTLR9 cells overexpress the human TLR9 (hTLR9) gene. They also express a SEAP (secreted embryonic alkaline phosphatase) reporter gene under the control of an NF-κB inducible promoter comprised of an IFN-β minimal promoter fused to five NF-κB and AP-1 binding sites. Levels of SEAP in the supernatant can be easily determined with HEK-Blue
Detection, a cell culture medium that allows real-time detection of SEAP activity.Unlike their parental cells, HEK-Blue
Null1, which weakly express TLR9, HEK-Blue
hTLR9 cells are responsive to stimulation with TLR9 agonists, such as oligonucleotides containing CpG motifs (CpG ODNs). As expected, hTLR9 overexpression in HEK-Blue
hTLR9 cells allows potent NF-κB responses upon incubation with human-preferred, but not mouse-preferred, CpG-ODNs of class A (e.g. ODN 2216) and class B (e.g. ODN 2006). Of note, HEK-Blue
hTLR9 cells are more responsive to the class B, human-preferred, ODN 2006. These cells also respond to ODN 2395, a human/mouse-preferred class C (see Figures). Background:
Toll-Like Receptor 9 (TLR9) is one of the most studied pattern recognition receptors (PRRs) for nucleic acids. It is an endosomal receptor that triggers NF-κB- and IRF-mediated pro-inflammatory responses upon the recognition of unmethylated cytosine-phosphorothioate-guanosine (CpG) forms of DNA [1-3]. TLR9 agonists can be mimicked by synthetic oligonucleotides containing CpG motifs (CpG ODNs) [1,3].
More detailsKey features:
Verified overexpression of TLR9 gene by RT-PCR
Functionally validated with a selection of TLR9 agonists
Readily assessable SEAP reporter activity
Applications:
Study of the NF-κB-dependent TLR9 signaling pathway
Screening of novel specific activators or inhibitors of the TLR9 signaling pathway
References1. Kumagai Y. et al., 2008. TLR9 as a key receptor of the recognition of DNA. Adv. Drug. Deliv. Rev. 60(7):795-804.
2. Heinz L.X. et al., 2021. TASL is the SLC15A4-associated adaptor for IRF5 activation by TLR7-9. Nature. 581(7808):316-322.
3. Kayraklioglu N. et al., 2021. CpG oligonucleotides as vaccine adjuvants. DNA Vaccines: Methods and Protocols. Methods in Molecular Biology. Vol. 2197. p51-77.