Generating tools for functional genomics in C. elegans – Elegineer

The nematode Caenorhabditis elegans is a widely appreciated, powerful system in which to study fundamental biological mechanisms ultimately related to human health. We propose to develop novel genomic tools to manipulate the genome of C. elegans. First, we plan to generate new resources and new techniques for engineering chromosomal loci by homologous recombination. Second, we will attempt to mobilize engineered transposons in order to develop genomic tools such as gene trap systems that are still missing in the C. elegans genetic toolbox. This should facilitate our analysis of animal development and physiology and improve the modeling of human pathologies in this model system.
The project will be conducted by three teams:
- the group of Jean-Louis Bessereau (Institut de Biologie de l'Ecole Normale Supérieure, Paris) pioneered the use of the drosophila transposon Mos1 as a genetic tool in C elegans. They optimized insertional mutagenesis based on Mos1 mobilization in the germline . Recently, this group developed the first efficient technique, called MosTIC, to engineer C. elegans loci by homologous recombination.
- the group of Manfred Schmidt (DKFZ, Heidelberg) developed cutting-edge technologies to identify the insertion points of transposable elements in genomes. Specifically, they have previously developed a new integration site analysis method, linear amplification-mediated (LAM) PCR, addressing unknown DNA flanking sequences down to the single cell level from highly complex samples.
- the group of Corinne Augé-Gouillou (GICC, Tours) is a specialist of the molecular mechanics of Mos1 transposition. They recently engineered hyperactive versions of the Mos1 transposon and transposase.
In addition, we will interact with a French biotech company, which developed customized meganucleases for gene engineering, in order to implement the use of their reagents in C. elegans.

By combining the skills of these different teams, we should be able to:
- characterize a library containing about 80,000 Mos1 insertions and greatly expand the possibility to engineer C. elegans genes by the MosTIC technique.
- generate a hyperactive Mos1 system to improve the efficiency of MosTIC and be able to mobilize recombinant Mos1-based vectors
- use meganucleases for gene transfer in C. elegans and develop efficient protocols for single copy transgenesis and targeting of C. elegans chromosomal loci.