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Boston Univeristy bioengineers devise 'dimmer swith' to regulate gene expression in mammal cells

messenger RNA (mRNA) -- from being made. If any mRNA gets past this repressor, the second technique, interfering RNA (RNAi) attaches to the functional mRNA, rendering it useless. The cell cannot turn it into protein.

I was delighted to see that when the two systems are coupled, it is possible to completely turn a genes function off, said Deans.

This switch is also reversible and tunable. By adding a chemical -- Isopropyl--thiogalactopyranoside -- the repressor components are blocked and the gene turns on again. The genes activity can be tuned up or down by adjusting the amount of this chemical.

The researchers demonstrated the strength of their off switch by hooking it up to the gene for diphtheria toxin, then inserting it into cells. One molecule of diphtheria toxin can kill a cell, but with the genetic switch turned off, the cells survived for weeks. When the researchers flipped the switch, toxin production was triggered and the cell died.

They also showcased the switchs capability for delicately tuning gene expression, by installing it alongside a gene that leads to apoptosis, programmed cell death, once a certain threshold concentration of the genes product is reached. They gradually increased the genes activity until they met and passed this threshold.

This tuning feature is important, said Deans, because many diseases are not a result of missing a gene, but rather a result of too much or too little expression. With the ability to tune the level of gene expression in our switch, we could explore threshold responses and how these result in disease phenotypes.

The switch may also hold promise for therapeutic applications. It gives a really nice regulator scheme for cell and gene therapy, said Collins. I think in the coming decades well increasingly see these therapies being introduced as part of routine medical practice.


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Contact: Mike Seele
mseele@bu.edu
617-353-9766
Boston University
26-Jul-2007


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