The consortium and project resources

We have assembled a team that is uniquely qualified to carry out the proposed research and to achieve the objectives within the outlined timeframe. One partner Jean-Louis Bessereau (Partner #2) was the first to achieve the controllable mobilization of a Drosophila transposon in the C. elegans germline (Bessereau et al, 2001, Nature, 413, 70-74). Since starting his own group in Paris, and with the aid of an INSERM-funded collaboration with the group of Jonathan Ewbank (Partner #3), he has continued to improve the system in order to generate an efficient tool for forward genetic screens. Specifically, his group has cloned five new genes involved in nicotinic neurotransmission using Mos1-mediated mutagenesis, while the group of Jonathan Ewbank has already applied it successfully to the dissection of innate immune responses in C. elegans. In addition to such forward genetic screens, the group of Laurent Segalat (Partner #6) has been running a pilot experiment to assay the feasibility of using the Mos1 system as a tool for generating a comprehensive collection of insertions that would tag most of the >19,000 C. elegans genes. This experiment is nearing completion and the generated data set of approximately 1000 characterized Mos1 insertions will be sufficiently large to confirm robustly preliminary results that indicate that Mos1 will be a suitable tool for whole genome reverse genetic screens in terms of an absence of significant insertion site bias, and the stability and intrinsic mutagenicity of Mos1 insertions.

Future efficient and practical directed and non-directed alterations of the C. elegans genome will require the use of transposons capable of carrying exogeneous DNA, such as fluorescent marker genes. A fundamental limitation to the Mos1 transposon is its limited DNA-carrying capacity. The co-ordinator of the project, Nektarios Tavernarakis, together with Partner #6, will develop alternative transposon-based tools to address this need. We have joined forces with MAIA Scientific, a company specializing in platform technology development, which will provide solutions essential for the automation and streamlining of the processes involved in generating libraries of mutant strains.  MAIA Scientific is a Harvard Bioscience Operating Company (NASDAQ: HBIO).  MAIA Scientific invents, develops, builds and delivers novel instrumentation for high throughput cell biology, genomics and drug discovery. Using instrumentation know-how, the company develops dedicated assay technology for customers and provides access to technology through contract R&D, partnerships and collaboration.  MAIA Scientific has already installed its automated worm sorter in two of the participating labs (# 3 and that of Patty Kuwabara, Partner # 5), that are both actively involved in platform development.

            The automated generation of a bank of mutants and the development of new transposon tools are but the first steps in a much broader functional genomic analysis of the whole C. elegans genome. This larger project will require the participation of a much broader constituency of qualified laboratories. One aspect of such a future project will be the capacity to generate transcriptional profiles for large numbers of mutants. Two laboratories (#3 and #5) are involved in the development of community-oriented microarray facilities, which are otherwise only available in the USA.

Means of optimal utilization/mobilization of resources

We will achieve optimal utilization and mobilization of resources by adhering to the following principles:

1.    Joint production and exchange of research materials - e.g. antibodies, clones, constructs, strains, etc. produced by one group will be available to other groups when they need them.

2.    Joint experiments - e.g. shared subprojects, in which part of the work is done by one group and another part by the other group(s).

3.    Collaboration through training - e.g. transfer of specific skills (theoretical and practical), or techniques used routinely by one group, will be taught to members of another group in short term visits.

4.    Exchange of information - e.g. new knowledge relevant to the successful execution of tasks, produced by one group will be disseminated to other groups.

5.    Sharing infrastructure - instruments available in one organization will be used by members of other organizations in short term visits.

All the equipments necessary for the execution of the project are already available in the individual laboratories and will be used by the consortium researchers as needed.

List of the main facilities, specific equipments that will be integrated:

-    Computing facilities

-    Microinjection equipment

-    Collection of C. elegans stocks.

-    Protein purification facilities (FPLC, HPLC systems, SMART micro-purification system)

-    DNA synthesis and sequencing facilities

-    Confocal microscopes (Leica TCS SP, Leica TCS-NT, Zeiss Axioplan 2 outfitted with a BioRad Radiance 2100 scan-head and laser resources).