THP1-Dual™ KO-IRF3 Cells
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Cat.code:
thpd-koirf3
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ABOUT
IRF3 knockout dual reporter monocytes
THP1-Dual™ KO-IRF3 cells were generated from the THP1-Dual™ cell line, which is derived from the human THP-1 monocytes, through the stable knockout of the IRF3 gene. IRF3 (interferon regulatory factor 3) is a critical player in the innate immune responses to virus infections. Viral nucleic acids in the cytosol are recognized by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs), RIG-I-like receptors (RLRs), or cytosolic DNA sensors (e.g. the cyclic GMP-AMP synthase cGAS). Upon ligand binding, these receptors trigger the production of type I interferons (IFNs) through the activation of the TBK1/IKKε-IRF3/IRF7 pathway [1]. Activated PRRs also trigger the production of pro-inflammatory cytokines through activation of the NF-κB pathway [1].
THP1-Dual™ KO-IRF3 and THP1-Dual™ 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.
As expected, the responses of THP1-Dual™ KO-IRF3 cells to the cyclic dinucleotide 2'3'-cGAMP (a STING agonist) and to lipopolysaccharide (a TLR4 agonist) are impaired. However, differential responses (either no effect, decrease or increase) are observed when using RNA or DNA agonists coupled with transfection reagents. THP1-Dual™ KO-IRF3 cells retain the ability to respond to cytokines such as type I IFNs and TNF-α. These cells are resistant to Blasticidin and Zeocin™.
Features of THP1-Dual™ KO-IRF3 cells:
- Biallelic knockout of the IRF3 gene
- Readily assessable Lucia luciferase and SEAP reporter activity
Applications for THP1-Dual™ KO-IRF3 cells:
- Defining the role of IRF3 in PRR-induced signaling
- Highlighting possible overlapping PRR activation or regulatory mechanisms (see 'Details')
Validation of THP1-Dual™ KO-IRF3 cells:
- IRF3 knockout verified by PCR, DNA sequencing, and western-blot
- Functionally tested
References:
1. Iurescia S. et al., 2018. Nucleic acid sensing machinery: targeting innate immune system for cancer therapy. Recent Pat. Anticancer Drug Discov. 13: 2-17
Disclaimer: These cells are for internal research use only and are covered by a Limited Use License (See Terms and Conditions). Additional rights may be available.
SPECIFICATIONS
Specifications
Defining the role of IRF3 in PRR-induced signaling, Highlighting possible overlapping PRR activation or regulatory mechanisms
Complete RPMI 1640 (see TDS)
Verified using PlasmotestTM
Each lot is functionally tested and validated.
CONTENTS
Contents
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Product:THP1-Dual™ KO-IRF3 Cells
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Cat code:thpd-koirf3
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Quantity:3-7 x 10^6 cells
- 1 ml of Normocin™ (50 mg/ml)
- 1 ml of Zeocin® (100 mg/ml)
- 1 ml of Blasticidin (10 mg/ml)
- 1 tube of QUANTI-Luc™ 4 Reagent, (Lucia luciferase detection reagent)
- 1 ml of QB reagent and 1 ml of QB buffer (SEAP detection reagent)
Shipping & Storage
- Shipping method: Dry ice
- Liquid nitrogen vapor
- Upon receipt, store immediately in liquid nitrogen vapor. Do not store cell vials at -80°C.
Storage:
Caution:
Details
THP1-Dual™ KO-IRF3 cells have been functionally tested using various sources of nucleic acids (see the validation data document).
As expected, IRF and NF-κB responses are severely impaired when the THP1-Dual™ KO-IRF3 cells are incubated with STING agonists, such as 2’3’-cGAMP, which do not require transfection to access the cytosol. However, differential responses are observed when using RNA agonists with transfection reagents. The weak agonist 5’ppp-dsRNA loses its ability to induce an IRF response in KO-IRF3 cells when complexed with LyoVec™ or LTX. On the contrary, the IRF response is unexpectedly increased when using the highly potent agonist 3p-hpRNA complexed to LyoVec™ or LTX. Surprisingly, with either agonist, the NF-κB response is barely affected. These data suggest that the use of different agonists and transfection reagents could highlight overlapping RNA-sensing or regulatory mechanisms.
DOCUMENTS
Documents
Technical Data Sheet
Validation Data Sheet
Safety Data Sheet
Certificate of analysis
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