Tufts iGEM 2014
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Summaries
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Robust biofilm formation using a cyclic-di-GMP aptamer and investigating ethics and applications of engineered bactiophage
A long, noncoding massively expressed regulatory RNA (merRNA) discovered in Bdellovibrio
bacteriovorus is present in high levels during its dormant phase. The merRNA is believed to sequester
cyclic-di-GMP, much like a sponge. Since cyclic-di-GMP is a second messenger for various cellular
functions, including motility and biofilm formation, the Tufts iGEM team introduced this merRNA
sequence into E. coli. Constitutive expression of this merRNA transcript was shown to increase biofilm
formation. This property can be useful in microbe-based approaches to environmental remediation.
Earlier designs for phage delivery of the merRNA to disrupt biofilms inspired an investigation into the
policy surrounding engineered bacteriophage. Tufts iGEM will be convening a panel of experts from
various disciplines to put forth recommendations for the responsible use of phage in therapeutic and
industrial applications. A proposal will be drafted for a silk bandage containing a phage cocktail which
can prevent and treat infection by antibiotic-resistant bacteria.
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Ribosponge Project Description:
We have devised a method to introduce a DNA sequence which encodes an RNA aptamer (i.e. - an
oligonucleotide sequence which binds to a specific molecule) into a non-pathogenic E. coli strain. We
have dubbed this RNA aptamer a “ribosponge” due to its unique mode of action. The ribosponge binds
cyclic di-GMP, a secondary intracellular messenger which signals bacteria to enter a persistent or biofilm
state. The signal is universal among many species such as E. coli, P. aeruginosa, and M. tuberculosis.
Blocking the signal of c-di-GMP by binding it with an aptamer could prevent the persistent state in these
and other pathogens. In order to ferry the sequence encoding the aptamer from our non-pathogenic E.
coli into other bacteria of the same species, we plan on using an M13 phage which does not kill the
bacteria.
The project has also inspired our collaboration with the Rathneau Institute and SYNENERGENE as we
look at the feasability of developing ribosponge into a product, and examine the regulatory, legal, and
ethical challenges of packing it into a bacteriophage.
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Backgrounds
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Methods
Sponge Lab Notebook
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