Mouse Pipelines

Scientific Operations

The Mouse Pipelines team supports biomedical research through the humane and optimized use of genetically-modified mice. Under the most stringent care for the welfare of the animals, the team generates, phenotypes, cryopreserves, and distributes new mouse strains to galvanize biomedical research. This work supports the research efforts of the Sanger Institute Faculty, large international projects like EUCOMM, EUCOMMTools, Infrafrontier, Knock-Out Mouse Project, and Wellcome Trust Strategic Awards. The main technologies employed for the generation of the new strains are gene targeting in mouse embryonic stem cell (mESC) and CRISPR-mediated mutagenesis by zygote cytoplasmic microinjection. All the strains are made available to the scientific community worldwide as soon as they had passed appropriate quality control. Strains are preserved as frozen sperm and deposited at the main repositories in Europe and the US for long-term distribution. The effect of each mutation is also analyzed through a standard set of phenotyping tests and conditions. The phenotypic data is distributed to the scientific community through the portal of the International Mouse Phenotyping Consortium (IMPC).

The Mouse Pipelines aims to maximize the benefit of biomedical research using genetically modified mice through technological innovation, process optimization, high-quality management, and economies of scale, while maintaining a very high level of care for the welfare of the animals and the quality of the scientific data produced. Structurally, the team was divided into groups that naturally fitted the technical and experimental workflow.

The Genome Engineering group designs new alleles of biomedical importance to address the experimental objectives of the diverse projects we support. For CRISPR-mediated mutagenesis, the team prepares gRNAs and Cas9 mRNA for the Transgenic Technologies group to generate deletion alleles, and additionally single-stranded DNA to generate point mutations. For more complex alleles, like knockins or conditional alleles, gene targeting in mouse ES cells is employed.

The Transgenic Technologies group carries out blastocyst microinjection of mutant ES cells or cytoplasmic injection of CRISPR mutagenesis reagents into mouse zygotes to introduce the mutations into the mouse embryos. In coordination with staff of the Research Support Facility, the progeny derived from the microinjection experiment is bred to allow the transmission of the mutations into the germline of the F1 mice.

The Molecular Technologies group analysed DNA samples from the F1 mice to demonstrate that the desired allelic structure has been produced. This team also performs all the genotyping necessary to support the breeding of each mouse colony to produce all the cohorts necessary for phenotyping and other uses.

The Mouse Distribution group coordinates the export of strains. When the F1 mice have passed the quality control tests, they are adverstised as available to the scientific community. For the majority of the strains, two F1 heterozygote males are used to preserve the mutant allele for long term distribution in the form of frozen sperm. For some strains, frozen embryos are preserved instead of sperm. The frozen sperm or embryos are deposited at the European Mutant Mouse Archive (EMMA)/Infrafrontier, and at the United States KOMP and MMRRC repositories for long term storage and distribution. If requests are received while mice are still available from our internal breeding program we endeavor to provide the strains directly to any scientist in the world. The Mouse Pipelines works closely with staff of the Research Support Facility (RSF), with external requestors, and with European and American repositories to coordinate the export of mouse strains to the scientific community.

The Mouse Phenotyping group carries out a standardized battery of phenotypic tests applied to cohorts of 7 mutant males and 7 mutant females (+ matched wild type controls) for each of the mutant strains. All the tests are performed a specific ages and in a specific sequence to all the strains. The tests can be divided into 3 general categories: developmental, in vivo, and necropsy and blood analysis. The data is collected in database form, subjected to quality control, and submmited to the IMPC for exterizorization to the scientific community.

Historically, the Mouse Pipelines team has participated in several international large scale mutagenesis and phenotypic analysis. Included in this are the those funded by the European Union, like: EUMODIC, EUCOMM, EUCOMMTools, EMMAService, Infrafrontier-i3. We have also participated in the KnockOut Mouse Project Phases 1 and 2 funded by the US National Institutes of Health. Sanger Institute core funding has supported the Mouse Genetics Project. In addition the team is closely involved in three major Strategic Award projects funded by the Wellcome Trust and on the IMPC:

We work closely with the Mouse Informatics and Research Support Facility teams to develop the Mouse Database. This work is essential for the welfare of our mice, to maintain optimal operational workflows, and to achieve our scientific goals, as well as for streamlining publication of our scientific results through the International Mouse Phenotyping Consortium (IMPC) web portal.