Human cGAS-KO Dual Reporter THP-1 Cells

NF-κB-SEAP & IRF-Lucia reporter monocytes

ABOUT

cGAS knockout NF-κB-SEAP and IRF-Lucia luciferase reporter monocytes

THP1-Dual™ KO-cGAS cells were generated from THP1-Dual cells by stable knockout of the cGAS gene. They derive from human THP-1 monocytes, a cell line often used to study DNA-sensing pathways, as they express all the cytosolic DNA sensors identified so far (with the exception of DAI).

THP1-Dual™ and THP1-Dual™ KO-cGAS cells stably express two inducible secreted reporter genes: Lucia luciferase and SEAP (secreted embryonic alkaline phosphatase). The Lucia luciferase reporter gene is under the control of an ISG54 (interferon-stimulated gene) minimal promoter in conjunction with five IFN-stimulated response elements. The SEAP gene is driven by an IFN-β minimal promoter fused to five copies of the NF-kB response element. As a result, they allow the simultaneous study of the IFN regulatory factor (IRF) and the NF-kB pathway by assessing the activity of Lucia luciferase and SEAP, respectively. Both reporter proteins are readily measurable in the cell culture supernatant when using QUANTI-Luc™ 4 Lucia/Gaussia, a Lucia and Gaussia luciferase detection reagent, and QUANTI-Blue™ Solution, a SEAP detection reagent.

THP1-Dual™ KO-cGAS and THP1-Dual™ cells can be used to study the role of cGAS by monitoring IRF-induced Lucia luciferase activity.

 

References:

1. Sun L. et al., 2013. Cyclic GMP-AMP synthase is a cytosolic DNA sensor that activates the type I interferon pathway. Science 339(6121):786-91.
2. Gao P. et al., 2013. Cyclic [G(2’,5’)pA(3’,5’)p] is the metazoan second messenger produced by DNA-activated cyclic GMP-AMP synthase. Cell. 153(5):1094-107.
3. Ablasser A. et al., 2013. cGAS produces a 2’-5’-linked cyclic dinucleotide second messenger that activates STING. Nature. 498(7454):380-4.

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

Target

cGAS

Target species

Human

Tested applications

Screening of PRR agonists or inhibitors

Cell type
Monocytic
Growth properties
Suspension
Tissue origin
Human monocytes
Reporter gene
SEAP
Lucia®
Detection method
Colorimetric (SEAP), Bioluminescence (Lucia)
Antibiotic resistance
Blasticidin
Zeocin®
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-cGAS Cells
  • Cat code: 
    thpd-kocgas
  • Quantity: 
    3-7 x 10^6 cells
Includes:
  • 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 (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)

Shipping & Storage

  • Shipping method:  Dry ice
  • Storage:

    • Liquid nitrogen vapor
    Stability: 20 passages

    Caution:

    • Upon receipt, store immediately in liquid nitrogen vapor. Do not store cell vials at -80°C.

Details

THP1 reporter cells are a family of cells derived from the human monocytic THP-1 cell line, which naturally expresses many pathogen recognition receptors (PRRs), including Toll-like receptors.

They respond to ligands for certain TLRs; namely, TLR2, TLR1/2, TLR2/6, TLR4, TLR5 and TLR8. These cells can also be used to study DNA sensing pathways, as they are highly responsive to PRR agonists that trigger interferon (IFN) signaling pathways.

THP1‑Dual™ cells feature two reporter genes that enable the simultaneous study of the NF-κB and IFN signaling pathways.

Cyclic GMP-AMP synthase (cGAS, cGAMP synthase) is a critical cytosolic DNA sensor that triggers innate immune responses through the production of type I interferons (IFNs) [1]. In response to cytosolic double‑stranded DNA (dsDNA), cGAS produces the cyclic dinucleotide (CDN) 2’3’-cGAMP. CDNs bind directly to STING, leading to TBK1‑IRF3-mediated activation of IFN-stimulated response elements (ISRE) in the promoters of IFN-stimulated genes (ISG). The most potent agonist of human STING is 2’3’-cGAMP [2,3].

DOCUMENTS

Documents

THP1-Dual KO-cGAS Cells

Technical Data Sheet

Safety Data Sheet

Validation Data Sheet

Certificate of analysis

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