The EUCOMM group executes gene trapping and gene targeting mutagenesis of genes in ES cells under the auspices of EUCOMM and EUCOMMtools (The European Mouse Mutagenesis Consortium) programs. Gene targeting and high-throughput genotyping are integrated into a technologically advanced and robust production pipeline for this purpose

The state-of-the-art targeting pipeline is divided into two core areas, ES cell mutagenesis and high-throughput genotyping. Together, these two teams comprise a highly optimized targeting platform, mutating over 300 genes/month. EUCOMM (FP6) and EUCOMMtools (FP7) programs, which, together with independent programs KOMP/Regeneron (The Knockout Mouse Project), and NorCoMM (North American Conditional Mouse Mutagenesis Project) have been tasked with the targeted mutagenesis of approximately 20,000 protein coding genes, of which ~16,400 have already been completed. The alleles generated are conditional and amenable to further manipulation, creating an opportunity for wide-ranging functional studies. Thus, the next generation EUCOMMtools was initiated to develop 250 novel Cre-driver lines and a genetic toolkit for researchers seeking to fluorescently tag, purify, overexpress, or make toxic their locus of interest. Under the newly funded program, the group is responsible for the targeting, genotyping and archivation of the final 3,500 unique protein-coding genes in ES cells to be completed until 2015.

Since the inception of EUCOMM in 2006, the focus has been on attaining a highly reproducible pipeline for targeting and genotyping in ES cells. Overall, the throughput and success rate have increased sharply over the last five years, signifying productivity and QC gains. Several key accomplishments point to the success of the group:

  • Over 2963 genes have been correctly targeted in 3997 targeting attempts, which resulted in approximately 74% targeting efficiency.
  • Robust SOPs have been established for high-throughput gene targeting in ES cells and long-range PCR genotyping (6-9 kb).
  • A novel targeting vector cloning system has been developed using Saccharomyces cerevisiae as a host. The new system enables cloning of targeting vectors in less than a week; it is also amenable to high-throughput vector construction for genomic projects.

EUCOMM Success Rate vs. Vector Electroporations/Time

A JM8 ES cell colony ready for picking