Human RIG-I-KO Dual Reporter THP-1 Cells

NF-κB-SEAP & IRF-Lucia reporter monocytes

ABOUT

KO-RIG-I dual reporter monocytes for RLR pathway studies

InvivoGen offers a new series of THP1-Dual™ cell lines,  derived from the human THP-1 monocytic cell line, and specifically designed for assessing the role of RIG-I, MDA5, and MAVS in the cytosolic RNA sensing pathways. 

THP1-Dual™ KO-RIG-I cells express 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. In addition, these cells feature a stable knockout of the RIG-I (Retinoic Acid Inducible protein 1) gene.

 

RIG-I and MDA5 are two distinct sensors of viral double-stranded RNA (dsRNA), a replication intermediate for RNA viruses [1-3]. Upon recognition of dsRNA, RIG-I and MDA5 are recruited by the MAVS adaptor to the outer membrane of the mitochondria leading to the activation of several transcription factors, including interferon regulatory factors (IRFs) and NF-κB [3].

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Key features

  • Verified knockout of the RIG-I gene (PCR, DNA sequencing, Western blot, and functional assays)
  • Functionally validated with a selection of PRR ligands and cytokines
  • Readily assessable Lucia luciferase and SEAP reporter activities
  • The stability for 20 passages, following thawing, has been verified
  • Guaranteed mycoplasma-free

Application

  • Defining the role of RIG-I in cytosolic RNA sensing pathways
  • Highlighting possible overlap between RIG-I/MDA5 and STING signaling functions
  • Developing novel specific inhibitors of the RLR signaling pathway

 

References

1. Gebhardt A. et al., 2017. Discrimination of Self and Non-Self Ribonucleic Acids. Journal of Interferon & Cytokine Research 37: 184-97.
2. Pichlmair A. et al., 2006. RIG-I mediated antiviral responses to single-stranded RNA bearing 5’-phosphates. Science 314:997-1001.
3. Kawai T. et al., 2005. IPS-1, an adaptor triggering RIG-I- and Mda5-mediated type I interferon induction. Nat Immunol. 6(10):981-988.

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.

Reporter systems in THP1-Dual™-derived cells
Reporter systems in THP1-Dual™-derived cells

SPECIFICATIONS

Specifications

Cell type
Monocytic
Growth properties
Suspension
Tissue origin
Human B cell lymphoma
Reporter gene
SEAP
Lucia®
Antibiotic resistance
Zeocin®
Blasticidin
Growth medium

Complete RPMI 1640 (see TDS)

Mycoplasma-free

Verified using Plasmotest

Quality control

Each lot is functionally tested and validated.

CONTENTS

Contents

  • Product: 
    THP1-Dual™ KO-RIG-I Cells
  • Cat code: 
    thpd-korigi
  • Quantity: 
    3-7 x 10^6 cells
Includes:
  • 1 ml of Blasticidin (10 mg/ml)
  • 1 ml of Zeocin® (100 mg/ml)
  • 1 ml of Normocin™ (50 mg/ml). Normocin™ is a formulation of three antibiotics active against mycoplasmas, bacteria, and fungi.
  • 1 ml of QB reagent and 1 ml of QB buffer (sufficient to prepare 100 ml of QUANTI-Blue™ Solution, a SEAP detection reagent)
  • 1 tube of QUANTI-Luc™ 4 Reagent, a Lucia luciferase detection reagent (sufficient to prepare 25 ml)

Shipping & Storage

  • Shipping method:  Dry ice

    • Storage:

    • Liquid nitrogen vapor
    Stability: 20 passages

Details

RIG-I (retinoic-acid-inducible protein 1, also known as Ddx58)

RIG-I is a cytoplasmic RNA helicase that is critical for host antiviral responses. It senses double-stranded RNA (dsRNA), a replication intermediate for RNA viruses, leading to the production of type I interferons (IFNs) [1]. RIG-I binds specifically to short dsRNAs that have blunt ends and a 5’-triphosphate (5’-ppp) moiety, facilitating discrimination between host and viral dsRNA [2].

MDA5 (melanoma-differentiation-associated gene 5, also known as Ifih1 or Helicard)

MDA5 is a cytoplasmic RNA helicase that plays an important role in antiviral response. It senses long double-stranded RNA (dsRNA), a replication intermediate for RNA viruses, leading to the production of type I interferons (IFNs) [1]. MDA-5 and the related RNA helicase RIG-I recognize a complementary set of cytosolic viral dsRNA. Transfected Poly(I:C), a synthetic analog of viral dsRNA, is recognized by both MDA-5 and RIG-I.

MAVS (mitochondrial antiviral-signaling protein, also known as IPS‑1, CARDIF, VISA)

MAVS is an adaptor protein that plays a critical role in the immune response to viral infection. The innate immune system senses intracellular double-stranded RNA (dsRNA), a replication intermediate for RNA viruses, through two RNA helicases: retinoic acid-inducible gene-I (RIG-I) and melanoma differentiation-association gene 5 (MDA5). Upon recognition of dsRNA, RIG-I and MDA5 are recruited by MAVS to the outer membrane of the mitochondria leading to the activation of several transcription factors including interferon-regulatory factor 3 (IRF3), IRF7, and NF-κB. IRFs and NF-κB regulates the expression of type I interferons (IFNs) and pro-inflammatory cytokines, respectively [1, 3].

 

1. Gebhardt A. et al., 2017. Discrimination of Self and Non-Self Ribonucleic Acids. Journal of Interferon & Cytokine Research 37: 184-97.
2. Pichlmair A. et al., 2006. RIG-I mediated antiviral responses to single-stranded RNA bearing 5’-phosphates. Science 314:997-1001.
3. Kawai T. et al., 2005. IPS-1, an adaptor triggering RIG-I- and Mda5-mediated type I interferon induction. Nat Immunol. 6(10):981-988.

DOCUMENTS

Documents

THP1-Dual™ KO-RIG-I Cells

Technical Data Sheet

Validation Data Sheet

Safety Data Sheet

Certificate of analysis

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Citations

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