THP1-Dual™ KO-TREX1 Cells

TREX1 knockout NF-κB-SEAP and IRF-Lucia luciferase reporter monocytes

THP1-Dual™ KO-TREX1 cells were generated from THP1-Dual™ cells by stable knockout of the TREX1 gene. TREX1 (also known as DNAse III) is a major cellular 3'5' exonuclease that plays a crucial role in maintaining immune homeostasis [1,2]. These cell lines were derived 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-TREX1 cells stably express two inducible secreted reporter genes: IFN-inducible Lucia luciferase and NF‑κB-inducible SEAP (secreted embryonic alkaline phosphatase). They can be used to study the role of TREX1 by monitoring IRF-induced Lucia luciferase activity, using QUANTI‑Luc™, a Lucia™ luciferase detection reagent.

THP1-Dual™ KO-TREX1 cells are resistant to the selectable markers blasticidin and Zeocin®.

References:

1. Kavanagh D. et al., 2008. New roles for the major human 3'-5' exonuclease TREX1 in human disease. Cell Cycle. 7(12):1718-25.
2. Hasan M. & Yan N., 2014. Safeguard against DNA sensing: the role of TREX1 in HIV-1 infection and autoimmune diseases. Front Microbiol. 5:193

Figures

(A) The targeted TREX1 region in THP1-Dual™ (WT) and THP1-Dual™ KO-TREX1(KO) cells was amplified by PCR. THP1-Dual™ KO-TREX1 cells feature a biallelic deletion (arrow). MWM = molecular weight marker. (B) Lysates from THP1-Dual™ (WT) and THP1-Dual™ KO-TREX1 (KO) cells were analyzed by Western blot (Wes™) using an anti-human TREX1 antibody, followed by an HRP conjugated anti-rabbit secondary antibody. The arrow indicates the expected band for the TREX1 protein (33 kDa).

Specifications

Antibiotic resistance: Zeocin®, blasticidin

Growth medium: RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum, 100 μg/ml Normocin™, Pen-Strep (100 U/ml-100 μg/ml)

Quality control
TREX1 knockout is verified by PCR and DNA sequencing.
These cells are guaranteed mycoplasma-free.

This product is covered by a Limited Use License (See Terms and Conditions).

Contents

• 1 vial of THP1-Dual™ KO-TREX1 cells (3-7 x 10⁶ cells) in freezing medium
• 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 pouch of QUANTI-Luc™
• 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)

Description

THP1-Dual™ and THP1-Dual™ KO-TREX1 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‑κB response element. As a result, they allow the simultaneous study of the IFN regulatory factor (IRF) pathway, by assessing the activity of Lucia luciferase and the NF-κB pathway, by monitoring the activity of SEAP. Both reporter proteins are readily measurable in the cell culture supernatant when using QUANTI‑Luc™, a Lucia™ luciferase detection reagent and QUANTI-Blue™, a SEAP detection reagent.

THP1-Dual™ KO-TREX1 and THP1-Dual™ cells can be used to study the role of TREX1 by monitoring IRF-induced Lucia luciferase activity.