|HEK-Blue™ mTLR13||Unit size||Cat. code||Docs||Qty||Price|
murine TLR13-expressing HEK 293 cells
3-7 x 10e6 cells
Mouse TLR13 reporter cell line
HEK-Blue mTLR13 cells are HEK293-derived cells stably transfected with pUNO-mTLR13 and an NF-κB-inducible SEAP (secreted embryonic alkaline phosphatase) reporter plasmid.
Stimulation with a TLR13 ligand, such as ORN Sa19, activates NF-kB and leads to the production of SEAP. Levels of SEAP can be easily determined with HEK-Blue™ Detection, a growth & detection medium that turns purple/blue in the presence of alkaline phosphatase.
The parental cell line of HEK-Blue™ mTLR13 cells is HEK-Blue™-Null1-v cells.
Growth medium: Growth Medium: DMEM, 4.5 g/l glucose, 10% (v/v) fetal bovine serum, 50 U/ml penicillin, 50 µg/ml streptomycin, 100 µg/ml Normocin™, 2 mM L-glutamine.
Guaranteed mycoplasma-free.Back to the top
- 1 vial of HEK-Blue™-mTLR13 Cells (3-7 x 10e6 cells) in Freezing Medium
- 100 μl Blasticidin selective antibiotic (10 mg/ml).
- 100 μl Zeocin™ selective antibiotic (100 mg/ml).
- 1 ml Normocin™ (50 mg/ml).
- 1 pouch of QUANTI-Blue™, alkaline phosphatase detection medium.
Shipped on dry iceBack to the top
HEK-Blue™-mTLR13 cells are designed for studying the stimulation of mouse TLR13 (mTLR13) by monitoring the activation of NF-κB.
HEK-Blue™-mTLR13 cells were obtained by co-transfection of the mTLR13 gene and an optimized secreted embryonic alkaline phosphatase (SEAP) reporter gene into HEK293 cells. The SEAP reporter gene is placed under the control of the IFN-β minimal promoter fused to five NF-κB and AP-1 binding sites. Stimulation with a TLR13 ligand activates NF-κB and AP-1 which induces the production of SEAP.
Levels of SEAP can be easily determined with QUANTI-Blue™ a detection medium that turns purple/blue in the presence of alkaline phosphatase.
HEK293 cells express endogenous levels of TLR3, TLR5 and NOD1.
Expression of the mouse TLR13 gene was confirmed by RT-PCR.
HEK-Blue™-mTLR13 Cells were stimulated by the TLR13 agonist ORN Sa19. As expected, TLR13 agonists induced the production of SEAP.
These cells are guaranteed mycoplasma-free.
Specificity of HEK-Blue™-mTLR13 Cells
As HEK293 cells express endogenous levels of TLR3, TLR5 and NOD1, HEK-Blue™-mTLR13 Cells will respond to their cognate ligands, such as poly(I:C), flagellin and iE-DAP, respectively. In order to identify TLR13-specific responses, we recommend to use HEK-Blue™-Null1-v Cells as a control cell line.
Note: HEK-Blue™-mTLR13 Cells may be stimulated in a TLR13-independent manner as NF-kB/AP-1 can be activated by a wide variety of stimuli
Toll Like Receptor 13
A receptor for bacterial ribosomal RNA
Toll-like receptor 13 (TLR13) is an endosomal TLR expressed in mice, which role and ligand remain unclear.
Three separate groups have identified 23S ribosomal RNA (rRNA) as a ligand for TLR13. This single-stranded rRNA is present in gram-positive and gram-negative bacteria but not in eukaryotic cells. A conserved sequence of 10 residues within the catalytic center of 23S rRNA, “CGGAAAGACC”, was found to be both necessary and sufficient to trigger TLR13 signaling.
Other forms of rRNA are unable to activate the TLR13 pathway. Thus, unlike other nucleic acid receptors, TLR13 appears to recognize a specific RNA sequence.
Interestingly, Oldenburg et al. have shown that this sequence is the binding site of the macrolide- lincosamide-streptogramin (MLS) group antibiotics, and that methylation-mediated resistance to these antibiotics (which target
A2085 in S. aureus 23S rRNA) abolishes the immunostimulatory activity of 23S rRNA. This study suggests that acquisition of antibiotic resistance is a mechanism developed by bacteria to evade host innate immune system.
Humans lack TLR13 and probably rely on other pathogen receptors to detect pathogenic bacterial infection. The TLR13 signaling cascade clearly engages MyD88 and UNC93B1, but the details of the pathway require further investigation.
1. Oldenburg M. et al., 2012. TLR13 recognizes bacterial 23S rRNA devoid of erythromycin resistance-forming modification. Science. 337(6098).
2. Hidmark A. et al., 2012. Cutting edge: TLR13 is a receptor for bacterial RNA. J Immunol. 189(6):2717-21.
3. Li XD & Chen ZJ. 2012. Sequence specific detection of bacterial 23S ribosomal RNA by TLR13. elife. 1:e00102.