Targeted conditional somatic mutagenesis defines an entirely new scientific field, driven by internationally coordinated initiatives (EUCOMM, KOMP, NorCOMM) established for the systematic generation of conditional mouse mutants on a large scale.
The majority of these initiatives are committed to the production of mutant mouse ES cell lines, each of which carries an altered or "floxed" allele of a single gene. These mutant ES cell mutations can be readily transformed into mice using blastocyst injection, and the mutation activated by crossing the mouse bearing the floxed allele with a Cre recombinase driver strain to induce the mutation in spatially and temporally determined patterns.
Intricate conditional and inducible gene manipulation approaches have led to the generation of cell lineage- or developmental stage-specific alterations under temporal control. The vast number of Cre recombinase-expressing mouse lines - often referred to as "Cre-Zoo" animals - has greatly contributed to these accomplishments by allowing Cre recombinase to be expressed in specific cell types, in some cases in an inducible manner.
The full power of conditional mutant ES cell libraries and mice can therefore only be exploited with the availability of well characterised mouse lines expressing Cre-recombinase in tissue, organ and cell type-specific patterns, to allow the creation of somatic mutations in defined genes.
Although several privately curated and locally held databases currently provide a limited catalog of existing Cre driver strains, common problems are:
- These are not well integrated and often outdated, so that much of the field works by "word of mouth" to locate the necessary reagents for generating their conditional mutations.
- Published data on Cre driver mice from disparate groups do not always capture critical details such as the efficiency of recombination, cell and tissue specificity, or genetic background effects.
CREATE will address these combined shortcomings - inaccessibility to existing Cre driver strains, their incomplete characterisation, and an inadequate coverage of cell and tissue types in which they are active by enlisting major mouse stakeholders dedicated to collecting, integrating and curating, expanding and disseminating the currently scattered Cre driver mouse databases through a unified portal to provide the necessary structure for worldwide access to these critical resources.
Goals of CREATE
Disseminate extant information on Cre mouse resources through generation of an international CreZOO database with linkages that capture existing knowledge of Cre driver mouse genetic construction, background, expression pattern and activity according to a single set of standards.
Develop a roadmap for harnessing emerging technologies and methods for improving Cre-mediated recombination in vivo through targeted, intensive workshops and a dedicated Cre8 web portal.
Define the requirements of the mouse research community, by collecting information on new Cre driver expression patterns necessary for in-depth analysis of mutagenesis in different cell and tissue types through a CreExpress forum.
This Coordination Action will complement the major investment European Commission has recently made into Projects that make use of the mouse as a model organism (eg, EUCOMM, EUMODIC) and into database coordination as a mode of disseminating their results throughout the scientific community (CASIMIR). The international dimension of activities coordinated through CREATE will provide the opportunity for Europe to provide benchmarks for the rest of the world and ensure cross-talk between mutagenesis, phenotype and gene expression data.
The CREATE consortium represents a core of major European and international mouse database holders and research groups involved in conditional mutagenes.
|Principal Investigator||Affiliated Institute||Country||Description|
|Andras NAGY||Mt. Sinai Hospital||Canada||Hosts the oldest Cre database (Cre-X-Mice), a key player in NorCOMM, the Canadian mouse mutagenesis initiative|
|Ewan BIRNEY||EMBL-EBI||UK||European Bioinformatics Institute|
|Janan EPPIG||Jackson Laboratory||USA||Holds Cre strains for MGI and IMSR|
|Nadia ROSENTHAL||EMBL-MR||Italy||International mouse biology research center: EMBL-Monterotondo|
|Ramiro RAMIREZ-SOLIS||Wellcome Trust Sanger Institute||UK||Involvment in EUCOMM and EUMODIC, hosting major mouse mutagenesis and phenotyping initiatives|
|Steve Brown||MRC Mammalian Genetics Unit (MGU)||UK||The MGU is the leading partner in EUMORPHIA (European Union Mouse Research for Public Health and Industrial Applications), a pan-European project to improve our understanding of human physiology and disease by developing informative mouse models.|
|Vassilis AIDINIS||BSRC Fleming||Greece||Hosts largest SPF mouse facility in Greece|
|Wolfgang WURST||HMGU||Germany||Coordinates EUCOMM and German Gene Trap Consortium|
|Yann HÉRAULT||GIE-CERBM||France||Dedicated database (MouseCre) targeting a wide array of tissues|
In addition, CREATE has international advisors that will form the International Coordination Group (ICG), and will participate in CREATE-funded workshop.
|Principal Investigator||Affiliated Institute||Country|
|A. Francis STEWART||Technische Universitaet Dresden, BioInnovationZentrum||Germany|
|Björn LÖWENADLER||AstraZeneca R&D||Sweeden|
|Colin McKERLIE||Toronto Centre of Phenogenomics||Canada|
|Elizabeth SIMPSON||CMMT, University of British Columbia||Canada|
|Geoff HICKS||Mammalian Functional Genomics Centre||Canada|
|Hongkui Zeng||Allen Institute for Brain Science||USA|
|Martin HRABÈ DE ANGELIS||HMGU, German Mouse Clinic||Germany|
|Moira O'BRYAN||Monash Institute of Medical Research and Australian Phenomics Network||Australia|
|Nathaniel HEINTZ||Rockefeller University||USA|
The quality and effectiveness of the coordination mechanisms of CREATE are associated with a work plan which is divided into 6 workpackages (WP):
- To create a European virtual repository of genetically modified mice
expressing Cre recombinase under the control of cell-specific elements
- To capture and disseminate knowledge on existing Cre transgenic tools
- To facilitate choice of transgenic tools to take full advantage of the
conditional mutant libraries being produced by the IKMC
- To promote European collaboration towards unified mouse databases
- To create CRE8, the official website of the CREATE consortium
- To establish a BioMart-based web portal, displaying current repository information of 3+ international Cre databases in genomic context
- To create an open forum for the management of knowledge and information
- To assimilate CASIMIR reports, EUMODIC SOPs. EURExpress etc.
- To identify and catalogue existing and emerging technologies on the
creation of Cre driver mice
- To recommend/suggest standards on genetic modification, data
submission and phenotypic analysis of Cre expressing mice
- To link international Cre databases towards a 'one stop shop'
- To provide a global virtual repository of genetically modified mice
expressing Cre recombinase under the control of cell-specific promoters
- To promote international collaboration amongst Cre mouse makers and users
- To identify biological/medical disciplines and cell lineages/tissues for which there are insufficient or inefficient Cre-driver mice worldwide
- To identify cell specific promoters/enhancers or alternate transcripts that could drive cell/tissue-specific Cre expression
- To prepare for a large scale project for Cre driver strain production,characterization and dissemination
WP6 Management of workflow