TLR8 KO Dual Reporter THP-1 Cells
THP1-Dual™ KO-TLR8 Cells | Unit size | Cat. code | Docs | Qty | Price |
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TLR8 Knockout - Human THP-1 Reporter Monocytes |
3-7 x 10e6 cells |
thpd-kotlr8 |
You may also need : THP1-Dual™ Cells | View more associated products ▼
TLR8 knockout dual reporter monocytes
THP1-Dual™ KO-TLR8 cells were generated from the THP1-Dual™ cell line through the stable knockout of the TLR8 gene. They feature two inducible reporter genes, allowing the concomitant study of the IRF and NF-κB pathways, by monitoring the Lucia luciferase and SEAP (secreted embryonic alkaline phosphatase) activities, respectively. As expected, NF-κB- and IRF-mediated responses are abolished in THP1-Dual™ KO-TLR8 cells upon incubation with TLR8 and TLR7/8 agonists when compared to the THP1-Dual™ cells, with no notable difference for the other ligands tested (see Figures).
Toll-like receptor 8 (TLR8) is a crucial intracellular sensor involved in the detection of viral and bacterial pathogens [1,2]. Similar to its structural homolog TLR7, TLR8 is activated by single-stranded (ss)RNA, as well as synthetic imidazoquinoline compounds such as R848. TLR8 and its structural homolog, TLR7, have similar signaling pathways but display different ligand specificities and expression patterns [2]. TLR8 activation triggers NF-κB- and IRF-mediated production of pro-inflammatory cytokines and type-I IFNs, respectively.
NF-κB and IRF signaling pathways in THP1-Dual™ KO-TLR8 cells
Key Features:
- Verified knockout of the TLR8 gene (PCR, DNA sequencing, and Western blot)
- Functionally validated with a selection of PRR ligands and cytokines
- Readily assessable Lucia luciferase and SEAP reporter activities
Applications:
- Defining the role of TLR8 in PRR-induced signaling, or other cell signaling pathways
- Highlighting the possible overlap between TLR8 and other signaling pathways
- Screening specificity of novel TLR8 agonists for use in vaccine adjuvants
References
1. Moen, S.H. et al., 2019. Human Toll-like Receptor 8 (TLR8) is an Important Sensor of Pyogenic Bacteria and Is Attenuated by Cell Surface TLR Signaling. Front Immunol, 10: 1209.
2. Georg P. & Sander L.E., 2019. Innate sensors that regulate vaccine responses. Curr. Op. Immunol. 59:31
Specifications
Growth medium: RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 10% (v/v) fetal bovine serum (FBS), 100 U/ml penicillin, 100 µg/ml streptomycin, 100 µg/ml Normocin™
Antibiotic resistance: Blasticidin and Zeocin®
Quality Control:
- Biallelic TLR8 knockout has been verified by PCR, DNA sequencing, Western blot, and functional assays.
- The stability for 20 passages, following thawing, has been verified.
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These cells are guaranteed mycoplasma-free.
InvivoGen's products are covered by a Limited Use License (See Terms and Conditions).
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- 3-7 x 106 THP1-Dual™ KO-TLR8 cells in a cryovial or shipping flask
- 1 ml of Normocin™ (50 mg/ml). Normocin™ is a formulation of three antibiotics active against mycoplasmas, bacteria, and fungi.
- 1 ml of Zeocin® (100 mg/ml)
- 1 ml of Blasticidin (10 mg/ml)
- 1 tube of QUANTI-Luc™ 4 Reagent, a Lucia luciferase detection reagent (sufficient to prepare 25 ml)
- 1 ml of QB reagent and 1 ml of QB buffer (sufficient to prepare 100 ml of QUANTI-Blue™ Solution, a SEAP detection reagent)
Shipped on dry ice (Europe, USA, Canada, and some areas in Asia)
Details
TLR8 background
Toll-like receptor 8 (TLR8) is a crucial sensor involved in the detection of viral and bacterial pathogens within endosomes [1,2]. Similar to its structural homolog TLR7, TLR8 surveys phagocytic cargo for the presence of microbial single-stranded (ss) RNA. Despite having similar signaling pathways, TLR7 and TLR8 display different ligand specificities and expression patterns [2]. TLR7 is essentially expressed by plasmacytoid dendritic cells (pDCs), whereas TLR8 is highly expressed by myeloid cells [2]. This suggests that TLR7 and TLR8 have evolved to mediate distinct immune responses. TLR8 contains two distinct ligand interaction sites that facilitate its recognition of RNA degradation products and small synthetic nucleoside analogs such as resiquimod (R848) [2,3]. Ultimately, this interaction induces a conformational change and leads to the activation of NF-κB- and IRF-mediated production of pro-inflammatory cytokines (e.g. IL-1β and IL-12) and type I IFNs (IFN-β), respectively. There is growing evidence for TLR8 as a critical sensor in the essential humoral immune response needed to achieve optimal vaccine efficiencies with live attenuated vaccines [2, 4]. Therefore, there are ongoing efforts to develop more potent, specific, and less toxic TLR8 agonists for vaccine adjuvants.
References:
1. Moen, S.H. et al., 2019. Human Toll-like Receptor 8 (TLR8) is an Important Sensor of Pyogenic Bacteria and Is Attenuated by Cell Surface TLR Signaling. Front Immunol, 10: 1209.
2. Georg P. & Sander L.E., 2019. Innate sensors that regulate vaccine responses. Curr. Op. Immunol. 59:31
3. Tanji H. et al., 2015. Toll-like receptor 8 senses degradation products of single-stranded RNA. Nat. Struct. Mol. Biol. 22:109.
4. Ugolini M. et al., 2018. Recognition of microbial viability via TLR8 drives TFH cell differentiation and vaccine responses. Nat. Immunol. 19:386.