International Knockout Mouse Consortium

Knockout-first Mutant Alleles

EUCOMMTOOLS will create 3500 C57Bl/6 conditional mutant alleles for single-exon (or otherwise previously conditionally untargeted) protein-coding mouse genes. These alleles will be made predominantly by introducing an 'artificial intron', containing a standard EUCOMM promoter-driven targeting cassette, into the coding sequence of the single-exon gene.

Cre Resources

EUCOMMTOOLS will engineer 500 new Cre C57Bl/6 ES cell lines by a variety of approaches including Cre knock-ins into genes with useful expression patterns and site specific integration of Cre expressing Bac vectors by RMCE into defined loci such as Rosa26 or Hprt. The majority of the resource will be made with inducible forms of Cre recombinase such as CreERT2. Up to 250 lines of Cre driver mice on a pure C57Bl/6 background will be generated and the Cre expression patterns documented and annotated in day P7 and P56. These mice will form a matched Cre driver resource for C57Bl/6 mice produced from conditional IKMC resources.

Research, Technology and Complementary Reagents

EUCOMMTOOLS will develop novel technologies to add value, depth and flexibility to existing IKMC ES cell and mouse resources. Key areas include:

• Development of novel recombinase based regulatory switches
• Exploration of zinc-finger nuclease stimulated homologous recombination strategies in fertilized oocytes
• Development and validation of complementary modular vector reagents which enable the construction of new useful knock-in alleles such as fluorescent and other reporters, site specific recombinases, and mutant cDNAs. These novel alleles can be constructed either by re-utilizing existing IKMC modular vector resources or directly modifying existing targeted IKMC ES cell lines by RMCE.

Structure of the EUCOMMTOOLS project

Societal Benefits

Funding & Structure