Bioassays - GMO based bioassays in food safety
11 important questions on Bioassays - GMO based bioassays in food safety
What is the principe of the se of a genetically modified organism in a bioassay?
What are the advantages of a GMO bioassay?
- more specific
- higher sensitivity
- in vitro or in vivo
For what can you use a GMO bioassay?
- act as ligands for endogenous receptors (e.g. estrogen receptors)
- induce specific repair responses (e.g. SOS DNA damage response to genotoxic agents)
- Higher grades + faster learning
- Never study anything twice
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What are the different reporter gene assays?
- Aromatic hydrocarbon (Ah) receptor (doxin like compounds)
- Steroid hormone & other nuclear receptors
- Estrogen receptor (ER)
- Androgen receptor (AR)
- Thyroid hormone receptor (TR)
- Retinoic acid receptor (RAR)
- Progesteron receptor (PR)
- Glucocorticoid receptor (GR) - Genotoxic compounds (SOS/UMU gene response)
How does the DR CALUX assay go?
- Transcription factor (100 kDa) is activated by ligand binding
- High affinity for aromatic compounds with planar structure (dioxins and polycyclic aromatic hydrocarbons (PAHs)
- Ligand binding: transformation and in nucleus: heterodimer formation with Ah receptor nuclear translocator (ARNT)
- AhR and ARNT have basic helix-loophelix-DNA binding domain
- binding to DNA induces gene expression
How can you calculate the affinity of a compound?
TEF=EC50(reference)/EC50(comp.)
How does an ER CALUX work and for what is it used?
Upon ligand binding the ER complex is phosphorylated and binds to the ERE site of the DNA initiating gene expression; in the transfected cells of the ER CALUX also of luciferase reporter gene.
How doe you detect an agonist and a antagonist in e ER CALUX?
With an antagonist this is the other way around. However this only works when you rist added a known agonist.
What is a genotoxic bioassay with a GMO?
For what do they use genetically engineerd microorganisms?
- It is more specific
- High sensitivity
- Possible in vitro or in vivo.
Advantage of in vivo is that you get information about: Absorption, distribution, metabolism, excretion of the compounds
For waht do they use GMO zebra fish?
Zebra fish lights up if you add dioxins.
This can give you a dose responce curve.
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