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Part Standards

How ETHERNO works
ETHERNO is a flexible approach of in-vivo functional DNA nanostructure synthesis. It consists of three functional components:

1. BioBricks encoding nanostructure component strands

In each BioBrick, the component strand sequence is followed by an HTBS, a stemloop for binding of HIV reverse transcriptase and B0054, a strong terminator. For nanostructures composed of multiple DNA strands of different sequences, the BioBricks required follow the same basic structure as shown in Fig. 1. The component strand sequences can be conveniently changed by seamless cloning methods, such as described in Fig. 2.

2. HIV reverse transcriptase (HIVRT)*

HIVRT binds to the HTBS and initiate polymerization of DNA on the mRNA transcribed from the BioBrick. mRNA translation is therefore blocked.

3. Murine leukaemia virus reverse transcriptase (MLRT)*

HIVRT polymerizes the component strand slowly after binding to the HTBS. MLRT speeds up DNA polymerization and exerts RNAase H activity. MLRT does not bind to HTBS.

*Plasmids (pHIV_pTp66p51 and pMLRT) used in this project courtesy of Voigt Lab, Synthetic Biology Center, MIT.

Advantages
Instead of purchasing chemically-synthesizing DNA strands again and again, we harnessed E. coli's synthetic ability to produce single-stranded DNA for real life applications. There are several advantages of MIT system over chemically-synthesized DNA.

Firstly, production cost for intense DNA aptamer and origami research can be reduced in the long run. Once the correct sequence are acquired and inserted into E. coli genome, single-stranded DNA of interest can be produced efficiently and cheaply.

Secondly, since the genome of E. coli can be engineered, for example, with a lac operon, ETHERNO brings us closer to achieve regulated production of functional nanostructures inside cells.

Thirdly, ETHERNO confers E. coli endless possibilities of new functions, because of the flexibility of DNA nanostructures. Drug carriers, biosensors, nano-motors...you name it!

Part Table

<groupparts>iGEM18 Hong_Kong_HKU</groupparts>