TRIF KO Dual Reporter THP1 Cells
THP1-Dual™ KO-TRIF Cells | Unit size | Cat. code | Docs | Qty | Price |
---|---|---|---|---|---|
TRIF Knockout - Human THP-1 Reporter Monocytes |
3-7 x 10e6 cells |
thpd-kotrif |
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TRIF knockout dual reporter monocytes
THP1-Dual™ KO-TRIF cells were generated from the THP1-Dual™ cell line, which is derived from the human THP-1 monocytic cell line, through the stable knockout of the TRIF gene. THP1-Dual™ KO-TRIF cells feature two reporter genes allowing the simultaneous study of the IRF pathway, by monitoring the activity of an inducible secreted Lucia luciferase, and the NF-κB pathway by monitoring the activity of an inducible SEAP (secreted embryonic alkaline phosphatase). Lucia luciferase and SEAP activities are readily assessable in the supernatant using QUANTI-Luc™ 4 Lucia/Gaussia and QUANTI-Blue™ Solution detection reagents, respectively.
TIR-domain-containing adapter-inducing interferon-β (TRIF), also known as TIR-containing adaptor molecule-1 (TICAM-1), is a key adaptor molecule in endosomal TLR-dependent signaling cascades. Upon recognition of lipopolysaccharide (LPS) by endocytosed TLR4, TRIF is responsible for mediating activation of interferon regulatory factor 3 (IRF3) and the production of type I interferons (e.g. IFNβ) [1]. Additionally, TRIF has been shown to induce TLR4-dependent 'late' activation of NF-κB. Importantly, the endosomal TLR4-TRIF signaling cascade is independent of the cell surface TLR4-MyD88 signaling pathway [2]. For comparing these two distinct TLR4 signaling cascades, InvivoGen also offers THP1-Dual™ KO-MyD cells.
NF-κB and IRF signaling pathways in THP1-Dual™ KO-TRIF cells
Key Features:
- Verified knockout of the hTRIF gene
- Functionally validated on a selection of TLR4 and other PRR ligands and cytokines
- Distinct monitoring of NF-κB and IRF activation by assessing the SEAP and Lucia luciferase activities
Applications:
- Defining the role of TRIF in PRR-induced signaling (i.e. TLR4), or related cell signaling pathways
- Highlighting possible overlap between TRIF and other signaling pathways
References:
1. Ullah, M.O. et al. 2016. TRIF-dependent TLR signaling, its functions in host defense and inflammation, and its potential as a therapeutic target. J Leukoc Biol 100, 27-45.
2. Ciesielska, A. et al. 2020. TLR4 and CD14 trafficking and its influence on LPS-induced pro-inflammatory signaling. Cell Mol Life Sci.
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 TRIF knockout has been verified by PCR, DNA sequencing, Western blot, and functional assays.
- The stability for 20 passages, following thawing, has been verified.
- These cells are guaranteed mycoplasma-free.
This product is covered by a Limited Use License (See Terms and Conditions).
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- 3-7 x 106 THP1-Dual™ KO-TRIF 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
TRIF Background
TIR-domain-containing adapter-inducing interferon-β (TRIF), also known as TIR-containing adaptor molecule-1 (TICAM-1), is a key adaptor molecule in TLR-dependent signaling cascades. Specifically, TRIF is responsible for mediating TLR4-dependent, MyD88 independent signaling upon recognition of lipopolysaccharide (LPS), as well as, TLR3-dependent signaling upon recognition of double-stranded (ds)RNA [1]. Ultimately, TRIF recruitment activates interferon regulatory factor-3 (IRF3) which regulates both type I interferon (IFN) gene expression and a range of IFN-stimulated genes (ISG) [1]. TRIF plays a central role in a range of innate immune responses including anti-viral host defenses, cell death, and homeostasis [1].
Additionally, TRIF has been shown to induce TLR4-dependent late activation of NF-κB through recruitment and activation of TRAF6 or receptor-interacting serine/threonine protein kinases 1 and 3 (RIPK1 and RIPK3) [2,3]. In addition to the key role played by RIPK1 & RIPK3 in necroptosis, their activation in this context has been shown to lead to the acute expression of pro-inflammatory cytokines [2]. Importantly, TLR4-dependent TRIF activation is independent of its activation of the MyD88-dependent (at the cell surface) pathway, which is implicated in ‘early’ activation of NF-κB and a subsequent pro-inflammatory response [3].
References:
1. Ullah, M.O. et al. 2016. TRIF-dependent TLR signaling, its functions in host defense and inflammation, and its potential as a therapeutic target. J Leukoc Biol 100, 27-45.
2. Najjar, M. et al. 2016. RIPK1 and RIPK3 Kinases Promote Cell-Death-Independent Inflammation by Toll-like Receptor 4. Immunity 45(1), 46-59.
3. Ciesielska, A. et al. 2020. TLR4 and CD14 trafficking and its influence on LPS-induced pro-inflammatory signaling. Cell Mol Life Sci.