1.use of mice in biomedical research 2.komp: activity and progress to date 3.rationale and planning...
TRANSCRIPT
1. Use of mice in biomedical research2. KOMP: Activity and progress to date3. Rationale and planning for phenotyping4. Request for continued support
Presentation Overview
Mice in Biomedical Research• The laboratory mouse has been considered the premiere experimental model of human biology and
disease since 1902 when it was first used to demonstrate how genetic traits could be transferred from parents to offspring via classical or “Mendelian” inheritance in mammals.
• In just over a century, an impressive array of genetic tools, reagents and processes has been developed in the mouse, including:
– Homozygous inbred strains– Recombinant inbred and consomic strains– Transgenic and knockout methods– Monoclonal antibodies– iPS cells
• Evidence that mice have played a key role in biomedical research is provided by the fact that at least 18 Nobel prizes have been awarded for work done using the mouse. Most recently, the 2007 Nobel Prize was awarded to Mario Capecchi, Martin Evans, and Oliver Smithies for their discovery of the "principles for introducing specific gene modifications in mice by the use of embryonic stem (ES) cells.”
Funded KOMP ProjectsThe Completion of a Comprehensive Mouse Knockout Resource
($47 M - 5 years - funded 9/06)
CHORI-Sanger-UC DavisRegeneron Pharmaceuticals, Inc.
Development and Improvement of Inbred ES Cell Lines for Use in Generation of Mouse Mutants($1.9 M - 2 years - funded 9/06)
U PennsylvaniaSamuel Lunenfeld Research Institute
A Data Coordination Center for the Knockout Mouse Project (KOMP)($2.5M - 5 years - funded 9/06)
The Jackson Lab
The KOMP Repository($4.8M - 4 years - funded 8/07)
UC Davis – CHORI
Goals and Progress
CSD Regeneron EuCOMM NorCOMM TIGM Total
Goal (ES) 5,000 3,500 8,000 500 17,000
Vectors 4991 3116 4271 596
Targeted ES 3064 2038 2883 236 8,221
Mice 189 160 351 12 700
The KOMP Website
www.knockoutmouse.org
Details
Complete information on allele design, vector, genotyping and sequencing primers are provided to the end-user.
The KOMP Repository
www.komp.org
The repository offers vectors, KO ES cells, parental ES lines, and support services. Orders are processed using a standard “shopping cart.”
Orders
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WT-orderedmice-orderedcryo-recovery-orderedgermplasm-orderedGLT-orderedmicroinj-orderedpremium-ESC-pckg-orderedESC-ordered
KOMP Repository Activities
Orders have recently increased to about 100/month.
Over 3,000 users have created accounts and registered interest in ~2,500 genes.
Kent Lloyd, UCDavis
Rationale for KOMP2
• Supporting a broad phenotyping effort would provide the following advantages: – A single cohort of mice would go through multiple phenotyping assays, so the
cost of producing multiple cohorts in different laboratories for phenotyping would be eliminated.
– Each mutant mouse strain would be characterized for a broad set of phenotypes in a way that will allow direct comparisons and result in a more thorough description of gene function.
– Quality standards will be established and maintained, so the data will be of the highest reliability.
– The risk of not finding a phenotype will be greatly reduced.– Important, but unpublishable, negative results will be captured.
• 3 workshops: Rome in 2007, Bar Harbor and Toronto in 2008 to establish vision for an IMPC & discuss international, coordinated phenotyping efforts – agreed that the way forward is to develop a business plan
• Medical Research Council/Wellcome Trust workshops in Nov 2008 and Oct 2009 to engage UK scientific community
• NIH Phenotyping meeting, Bethesda October 2009 (survey)
• EC–funded EUMODIC (Helmholtz, Munich; ICS, Strasbourg, MRC Harwell, WTSI) project is now doing broad-based phenotyping of 500 mutant lines – completion 2011
Planning for next phase: Phenotyping
The International Mouse Phenotyping ConsortiumMembers:The Wellcome TrustThe Medical Research CouncilThe Wellcome Trust Sanger CenterMRC HarwellEuropean CommissionThe National Institutes of HealthGerman Mouse ClinicToronto Center for Phenogenomics
Pending members:Australia Phenomics NetworkCzech RepublicInstitut Clinique de la SourisNanjing UniversityUniversitat Autonoma de Barcelona
Current funding:EuMODIC - $7M/yr + matching funds, 150 strainsSanger Mouse Genetics Program - $9M/yr, 200 strainsAustralia Phenomics Network - $10.5M/yr
Phenotyping of KOMP CSD mice at Sanger
http://www.sanger.ac.uk/mouseportal/
Reproduction• Fertility
Developmental Biology• Viability
• Embryonic lacZ pattern• Viability at E14.5
• MRI at E14.5
Neurology• SHIRPA assessment
• Hot plate (pain)• Open field (anxiety)
• Stress-induced hyperthermia
• Brain anatomy
Musculo-skeletal• Grip strength
• X-rays (skeleton)• Bone mineral density• Bone mineral content
Infectious disease susceptibility• Salmonella (bacterial)• Citrobacter (bacterial)• Plasmodium (malaria)
• Flu (viral)
Metabolism• Weight gain curve
• Food intake and energy expenditure)
• Fat/lean tissue assessment by DEXA• Glucose tolerance test
• Serum Insulin• Blood Clinical Chemistry
Senses• Slit lamp (front of the eye)
• Eye fundus (retina)• Auditory brain response (hearing
circuits)
Immune system• Complete blood count• White blood cell profile
• Immunoglobulin subclass profile
Cardiovascular• Blood clinical Chemistry
• Heart weight (hypertrophy)• Heart histology
• Eye fundus (retinal vessels)
Cancer• Micronuclei analysis
Hair and skin• Hair growth rate• Hair follicle stem
cell
General • Visible malformations
• Adult organ lacZ expression• Necropsy
• Tissue banking for histology
Ramiro Ramirez-Solis, WTSI
Genentech/Lexicon Mouse Phenotype Project472 Mouse knockouts were broadly phenotyped
130 (27%) strains had 1 phenotype245 (52%) strains had 2-5 phenotypes
Andy Peterson, Genentech
Cohort breedingPhenotypingData UploadCohort breeding
PhenotypingData Upload
KOMP Repository
Mouse Production
ES cells
Mice/Embryos
DatabaseWeb server
Data
Data
Mouse Phenotyping Center(s)
Mice/Embryos
KOMP ES cells
EuCOMM ES cells
Mice/Embryos
MicroinjectionGermline TransmissionLacZ stainingLethality/FertilityCryopreservation
TrackingAnalysisDisplay
Cohort breedingPhenotypingData Upload
KOMP Phenotyping Scheme