Team:Queens-Canada/idea

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Objective

Our main goal for this summer was to build a toolkit for the nematode Caenorhabditis elegans for use as a synthetic biology chassis. Our focus was on finding and selecting viable promoters, protein domains, and other components for use by future teams.

The motivation for our project is simple enough: to date, there has been little interest in using complex eukaryotic and multicellular organisms in iGEM, and we feel that the advantages of multicellular organisms outweigh the challenges involved in working with them.

We really want to encourage other teams to embrace the potential benefits of working in a complicated organism like C. elegans, which has been extensively studied as a model organism for developmental biology and was the first multicellular organism sequenced. As such, it represents one of the best opportunities for iGEM-style synthetic biology to make inroads into higher forms of life.

R.''C. elegans''

One diagnostic tool which has received a lot of attention recently are the channelrhodopsins, which were isolated from the green algae Chlamydomonas reinhardtii, and serve as light-activated nonspecific cation channels which can cause nerve cells to depolarise in response to certain wavelengths of light. These have been used extensively in research biology to interrogate the roles of the neurons in C. elegans and other species, allowing researchers to, for example, cause nematodes to contract all of their muscles at once and seize up in response to specific wavelengths of light.1

Acting as counterpart to the channelrhodopsins is a channel called halorhodopsin, isolated from halophilic archaeans, which causes the cell to uptake chloride anions, and thus can be used to hyperpolarise a neuron and prevent it from firing. Importantly, the halorhodopsin excitation frequency is well-separated from the excitation frequency of channelrhodopsin-2, and so they can be used in the same organism, or even the same cell, without significant cross-talk.

By combining these proteins in a single transgenic organism, behind specifically-chosen promoters which target only certain neurons, we hope to make a "remote control" worm, which can be instructed to move either forward or backward by exposing it to the appropriate wavelengths of light.

Footnotes and Citations

1: Nagel G et al., 2003 Channelrhodopsin-2, a directly light-gated cation-selective membrane channel. PNAS 100(24). doi:10.1073/pnas.1936192100

2: Han X, Boyden ES, 2007 Multiple-Color Optical Activation, Silencing, and Desynchronization of Neural Activity, with Single-Spike Temporal Resolution. PLoS ONE 2(3): e299. doi:10.1371/journal.pone.0000299