THP1-Dual KO-cGAS Cells
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Cat.code:
thpd-kocgas
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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
cGAS
Human
Screening of PRR agonists or inhibitors
Complete RPMI 1640 (see TDS)
Verified using Plasmotest™
Each lot is functionally tested and validated.
CONTENTS
Contents
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Product:THP1-Dual KO-cGAS Cells
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Cat code:thpd-kocgas
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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 (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
- Liquid nitrogen vapor
- Upon receipt, store immediately in liquid nitrogen vapor. Do not store cell vials at -80°C.
Storage:
Caution:
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
Technical Data Sheet
Validation Data Sheet
Safety Data Sheet
Certificate of analysis
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