Mouse production and availability Debora Bogani, Janet Kenyan, Martin Fray Biological Resources Group Medical Research Council Harwell, UK Microinjection/ES cell lab/ archiving/distribution The MRC Mouse Networks (MMN) MRC Mouse Network are a forum to:  Choose the genes to KO • The Gene List  Encourage uptake of resources • Mice • Phenotypic Data  • • • Collaborative research Research Consortia Advise on primary phenotyping Perform in-depth phenotyping . .... . .... . ..... Other academics • Request and receive mouse lines • Assist with assay design • Request protocols • Request data MMN Consortium Accelerate Drug Discovery Disorders of Bone and Cartilage Cardiovascular disease Developmental Abnormalities Models of Disease and Obesity Mouse Eyes and Vision Research Tissue Remodelling and Fibrosis Malignant Haematopoiesis Ion Channel Consortium Liver Disease Neurodegenerative Neuromouse Kidney and Urogenital System Respiratory Disease Macrophage Biology Immune system Total for MMN Rest of the community N 16 Consortia • Nominated genes • Assist with assay design • Critique and analyse data • Request and receive mouse lines om in at M ed ut an ge As t lin nes sa es y m d ad D e ve at e a l en opm M en ou qui ry t se lin O ut es st se an n di ng t re qu es ts MRC Mouse Networks activity 56 177 13 50 ES cell culture – workflow • • • • • • >600 ES cell lines expanded/year Cells imported into quarantine lab and expanded/re-frozen ES cells are MAP tested (pathogens) Comprehensive QC before injection Recovered/expanded in clean lab and re-frozen Ridged 7 day/week rota Microinjection capacity • • • • • All blastocyst injections inside SPF barrier 4 x microinjection stations 4 x dissection/surgical stations 6 x technicians trained in MI/surgery Cross over with the archiving & ES cell teams Harwell gene list • • • • • >1600 genes listed Track activity for MMN consortia Dynamic to reflect the shifting landscape Real time updates on projects are circulated via forum Email alerts listing genes availability Gene Adcy1 Atp1a3 Kcnj11 Nlrc4 Cpn2 Eif2b3 Fcrl6 Gfap Gtf2e2 MGI number MGI:99677 MGI:88107 MGI:107501 MGI:3036243 MGI:1919006 MGI:1313286 MGI:3618339 MGI:95697 MGI:1915403 New gene nominations • New nominations can be made at any time: [email protected] • Multiple clones are imported from KOMP/EUCOMM(WTSI) • Strict QC • ~48% clone pass QC • ~50% genes result in GLT Trisomy (metacentric) Chr 1 Current progress • >820 clones injected so far for IMPC • >360 GLT lines available from Harwell, • WTSI strain distribution What if no IKMC clones are available? Interrogate IKMC resource No tm1a clones available Nominate for targeting: EUCOMMTools NLAC/NARlabs, Taiwan (20/year) CRISPR/Cas9 in development Production time scales • Each nominated gene passes through many stages before GLT is confirmed Gene nominations from the MMN Production centres expand clones Prepare single cell suspension Grow ES cells after antibiotic selection Transfer embryos to recipient Select blastocyst host with different coat colour Use coat colour to select GA pups Wait for litter to be born >18 months Chimaera mating, confirm GLT, Neo deletion Accessing mice and ex vivo tissues • • • • • • Searching the IMPC website: www.mousephenotype.org Web link takes you to repository holding the mice Alternative alleles/ex vivo tissues may be available Don’t forget to ask! Status of genes nominated for production is also visible Register your interest and get status updates Imports from other production centres • Majority of the MMN lines will be generated in the UK • Other IMPC partners have been assigned MMN genes • We are happy to help acquire mice from around the world International Mouse Strain Resource IMSR Independent labs TIGM RIKEN APF CARD CMMR MMHCC MMRRC EMMA Harwell Jax • >15,000 mouse strains at your finger tips: www.findmice.org European Mouse Mutant Archive (EMMA) • EMMA archives mice generated in Europe www.infrafrontier.eu • Harwell offers a free archiving service High demand from the MMN & others • 177 IMPC mouse lines exported from Harwell • Organs and tissues from 22 strains • 56 investigators from the MMN amongst the recipients • 8 to 10 export requests/week for Harwell mice Measuring impact • Measuring impact - essential but challenging • Repositories track distribution very well • Added value is extremely hard to quantify – grants/publications What do we send out? • Frozen embryos/sperm/live mice – tm1a and tm1b alleles generated for every strain • Conditional allele (tm1c) made to order • Defined tissues/organs harvested from mice on the shelf • MEFs made from homozygous KO mice • Wax embedded slides • Training e.g. Cryocourse (2-5th Mar, 2015) We are here to help: contact us on [email protected]