THP1-Dual™ KO-DNase2 Cells
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Cat.code:
thpd-kodnase2
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ABOUT
DNase2 knockout NF-κB-SEAP and IRF-Lucia luciferase reporter monocytes
THP1-Dual™ KO-DNase2 cells were generated from THP1-Dual™ cells by stable biallelic knockout of the DNASE2 gene. Human THP1 monocytes or derived macrophages are a common cellular model to study DNA sensing as they naturally express all cytosolic DNA sensors identified so far (except DAI). THP1-Dual™ KO-DNase2 cells feature 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.
DNase2 (Deoxyribonuclease 2) is a lysosomal endonuclease present in macrophages. It exerts an important function in clearing DNA from phagocytosed apoptotic cells and from maturating erythroblast nuclei [1,2].
Key features:
- Biallelic knockout of the DNASE2 gene
- Functionally validated with a selection of PRR ligands and cytokines
- Readily assessable Lucia luciferase and SEAP reporter activities
Applications:
- Study of IRF and NF-kB-dependent DNase2 signaling pathways
- Screening of interactions between DNase2 and other signaling protein
- Study the role of DNase2 in innate immunity
1. Kawane K. et al., 2014. DNA degradation and its defects. Cold Spring Harb Perspect Biol. Jun 2;6(6).
2. Crow Y.Z. & Stetson D.B., 2021. The type I interferonopathies: 10 years on. Nat Rev Immunol. Oct 20:1–13.
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
Study of IRF and NF-kB-dependent DNase2 signaling pathways, Screening of interactions between DNase2 and other signaling protein, Study the role of DNase2 in innate immunity
Complete RPMI (see TDS)
Verified using Plasmotest
Each lot is functionally tested and validated.
CONTENTS
Contents
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Product:THP1-Dual™ KO-DNase2 Cells
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Cat code:thpd-kodnase2
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Quantity:3-7 x 10^6 cells
- 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 tube of QUANTI-Luc™ 4 Reagent, a Lucia luciferase detection 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, a SEAP detection reagent)
Shipping & Storage
- Shipping method: Dry ice
- Liquid nitrogen vapor
Storage:
Details
DNase2 (alternatively known as DNase II, DNase2a, or Acid DNase) is an endonuclease critical in cellular health and development.
It is localized in the lysosome of macrophages and shows peak activity at an acidic pH [2]. This enzyme is involved in DNA metabolism: it digests exogenous DNA of apoptotic cells and DNA of nuclei expelled from erythroid precursors [1]. Like TREX1, another critical nuclease also known as DNase III, DNase2 helps to rapidly remove nucleic acid fragments to prevent cells from accumulating self-DNA in the cytosol and triggering autoimmunity [3].
Defects in DNase2 functions allow undigested DNA fragments to leak into the cytosol and trigger the STING-TBK1-IRF3 signaling pathway [1]. As a consequence, pro-inflammatory cytokines (e.g., IFN-β) are constitutively expressed, causing interferonopathies and other autoimmune disease phenotypes (e.g. severe anemia, arthritis) [2,4].
1. Rodero et al., 2017. Type I interferon-mediated autoinflammation due to DNase II deficiency. Nat Commun 8, 2176.
2. Keyel, 2017. Dnases in health and disease. Developmental Biology 429(1).
3. Lan, Y.Y. et al., 2014. “Dnase2a deficiency uncovers lysosomal clearance of damaged nuclear DNA via autophagy.” Cell reports vol. 9,1: 180-192.
4. Crow, Y.J. &Stetson, D.B. 2021. The type I interferonopathies: 10 years on. Nat Rev Immunol.
DOCUMENTS
Documents
Technical Data Sheet
Validation Data Sheet
Safety Data Sheet
Certificate of analysis
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