Biocontainment

BeeT is intended to be used outside the lab, in beehives, where it is in close contact with nature. We cannot be sure about the effect on existing ecosystems if BeeT would be released in the environment, which is why it is engineered to die if it leaves the beehive. Our first measure to achieve this is a light-induced kill switch that relies on the balance of a bacterial toxin and an antitoxin that are expressed simultaneously. In the dark beehive, the system is unaffected. In the presence of blue light, a component of sunlight, the balance is disturbed in favour of the bacterial toxin, killing the bacterium.

As an additional safety measure, BeeT relies on the presence of a synthetic amino acid that is to be applied to the beehive. In the presence of the synthetic amino acid, catalytically dead Cas9 (dCas9) is produced that has the synthetic amino acid incorporated, at the expense of catalytically active Cas9. If BeeT escapes from the hive, the synthetic amino acid is no longer available, and dCas9 can no longer be formed. Instead, catalytically active Cas9 is produced which cuts the BeeT genome as well as any heterologous DNA that is present, thereby killing the bacterium and preventing horizontal gene transfer.

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