Mouse TLR7 Reporter HEK 293 Cells

NF-κB-SEAP reporter cells

ABOUT

NF-κB–SEAP reporter HEK293 cells expressing mouse TLR7

HEK-Blue™ mTLR7 cells were engineered from the human embryonic kidney HEK293 cell line to study the mouse Toll-like receptor 7 (TLR7)-dependent NF-κB pathway. This important pattern recognition receptor (PRR) recognizes viral single-stranded RNA (ssRNA) structures and subsequently triggers NF-κB and IRF immune responses[1].

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Description

HEK-Blue™ mTLR7 cells express the murine TLR7 gene as well as an NF-κB/AP-1-inducible SEAP reporter gene. SEAP (secreted embryonic alkaline phosphatase) levels produced upon TLR7 stimulation can be readily determined by performing the assay in HEK-Blue™ Detection, a cell culture medium that allows for real-time detection of SEAP. Alternatively, SEAP activity may be monitored using QUANTI-Blue™, a SEAP detection reagent.

Due to the expression of mTLR7, HEK-Blue™ mTLR7 cells strongly and stably respond to TLR7-specific ligands (see figures).

Interestingly, while HEK-Blue™ hTLR7 cells do not respond to TLR8 agonists (e.g. TL8-506), HEK-Blue™ mTLR7 cells do respond to these ligands (see figures). This may be explained by the strong homology between mTLR7 and mTLR8, and the possibility that these murine TLRs, but not their human counterparts, have evolved to detect a broad overlapping range of ligands. 

As HEK293 cells express endogenous levels of TLR3, TLR5, and NOD1, HEK-Blue™ mTLR7 cells will respond to their cognate ligands (see figures).

 

Key Features

  • Verified expression of murine (m)TLR7
  • Strong and stable response to TLR7-specific ligands
  • Distinct monitoring of NF-κB by assessing the SEAP activities

Applications

  • Defining the distinct role of TLR7 in the NF-κB-dependent pathway
  • Comparing human and murine TLR7 responses using the HEK-Blue™ hTLR7 cell line
  • Screening for TLR7-specific agonists or inhibitors in comparison with their parental cell line HEK-Blue™ Null2-k

 

References:

1. Georg P. & Sander L.E., 2019. Innate sensors that regulate vaccine responses. Curr. Op. Immunol. 59:31.

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

Species
Mouse
Target

TLR7

Target species

Mouse

Tested applications

Detection and quantification of mouse TLR7 activity

Cell type
Epithelial
Growth properties
Adherent
Tissue origin
Human embryonic kidney cells
Reporter gene
SEAP
Detection method
Colorimetric
Antibiotic resistance
Blasticidin
Zeocin®
Growth medium

Complete DMEM (see TDS)

Mycoplasma-free

Verified using PlasmotestTM

Quality control

Each lot is functionally tested and validated.

CONTENTS

Contents

  • Product: 
    HEK-Blue™ mTLR7 Cells
  • Cat code: 
    hkb-mtlr7
  • Quantity: 
    3-7 x 10^6 cells
Includes:
  • 1 ml Blasticidin (10 mg/ml)
  • 1 ml Zeocin® (100 mg/ml)
  • 1 ml Normocin™ (50 mg/ml)
  • 1 pouch of HEK-Blue™ Detection (cell culture medium for real-time detection of SEAP)

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

Toll-Like Receptor 7

In humans, four Toll-Like Receptor (TLR) family members TLR3, TLR7, TLR8, and TLR9, mainly found in the endosome, are specialized in sensing viral-derived components. TLR7 and TLR8 recognize single-stranded (ss)RNA structures, such as viral ssRNA, miRNA, and various synthetic agonists [1]. Despite their similarities in PAMP (pathogen-associated molecular pattern) recognition, structure, and signaling partners, they highly differ in expression profiles and signaling responses, with TLR7 being more involved in the antiviral immune response and TLR8 mastering the production of proinflammatory cytokines [2]. TLR7 is mainly found in plasmacytoid dendritic cells (pDCs) and B cells, whereas TLR8 is highly expressed in monocytes, monocyte-derived DCs (mDCs), and macrophages [3].

TLR7 signaling

Upon viral infection, TLR7 translocates from the endoplasmic reticulum via the Golgi into the endosomes. Subsequently, it undergoes proteolytic cleavage and dimerization [1,3]. Once activated, it recruits the adaptor protein MyD88 to trigger IRF, AP-1, and NF-kB responses via TRAF6 (TNF receptor-associated factor 6) [1,3]. Depending on the stimulus and cell type, TLR7-mediated signaling induces IFN-α and IFN-regulated cytokines or T helper 17 (Th17) polarizing cytokines, such as interleukin (IL)-1β and IL-23 [4].

TLR7 therapeutic targeting

The involvement of nucleic acid-sensing mechanisms in the immune response against infections and other diseases makes them interesting targets for drug design [4]. TLR7 agonists are currently been tested as vaccine adjuvants and immunomodulatory therapeutics. They are extensively studied in the context of viral infection (e.g. SARS-CoV-2, Influenza, HIV), autoimmune (e.g. asthma, Lupus), and autoinflammatory diseases (e.g. cancer) [1-4]. Understanding the fundamental differences between these two related receptors could potentially be harnessed to discover novel drugs and improve vaccine efficacy/safety [4].

 

References:

1. Martínez-Espinoza I & Guerrero-Plata A. 2022. The Relevance of TLR8 in Viral Infections. Pathogens. 11(2):134.
2. Salvi V, et al., 2021. SARS-CoV-2-associated ssRNAs activate inflammation and immunity via TLR7/8. JCI Insight.;6(18):e150542.
3. Georg P. & Sander L.E., 2019. Innate sensors that regulate vaccine responses. Curr. Op. Immunol. 59:31.
4. de Marcken M, et al., 2019. TLR7 and TLR8 activate distinct pathways in monocytes during RNA virus infection. Sci Signal.;12(605):eaaw1347.  

DOCUMENTS

Documents

HEK-Blue™ mTLR7 Cells

Technical Data Sheet

Safety Data Sheet

Validation Data Sheet

Certificate of analysis

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