The Jackson Laboratory Knockout Mouse Production and Phenotyping Project (JAX KOMP2)

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PROJECT SUMMARY/ABSTRACT Mice and humans share approximately 20,000 genes. To date, little data exists for more than half of these genes and nearly one third have no functional annotation. Because of the high degree of similarity between the mouse and human gene set, genetic data generated in mice can often be extrapolated to human gene function. Mouse models of genes with common functionality between mice and men can lead to new models of human disease, which are useful for drug screening, preclinical studies and deeper understanding of biological and disease mechanism. The goal of the Knockout Mouse Phenotyping Program (KOMP2) is to generate lines of mice that carry knockouts (KOs) for a genome-wide collection of mouse genes and subject the mice to broad based phenotyping. The Jackson Laboratory (JAX) proposes to merge its currently funded KOMP2 Mouse Production and Cryopreservation (U42) and Knockout Mouse Phenotyping (U54) operations as part of RFA-RM-15-017 Limited Competition: Knockout Mouse Production and Phenotyping Project (UM1). JAX KOMP2 proposes to use cutting-edge and cost-effective Cas9 RNA-guided nuclease (Cas9 RGN, also called CRISPR/Cas9) technology to generate, breed, cryopreserve and phenotype1500 lines of mice during the project period. Continued effort will be made to improve the Cas9 RGN technology so as to reduce costs, increase targeting efficiency, and create more complex mutant alleles. Genes will be selected in coordination with our KOMP2 and IMPC partners, and will focus on those with poor annotation, genes that have significant community demand and integrate with other NIH-support programs, and genes predicted to function in select pathways. To guarantee ready access to the community, we will ship mice to outside investigators while they are alive on the shelf and deposit the lines into the Mouse Mutant Regional Resource Center (MMRRC) repositories for future use. Broad based phenotyping on juvenile animals up to 18 weeks of age will be performed on all 1500 lines of mice using International Mouse Phenotyping Consortium (IMPC)-required and JAX-specific protocols. We will assess body weight and composition, metabolic and physiological parameters, and behavioral and cognitive function. To detect age-dependent phenotypes, we will use the same pipeline to phenotype 20% (300 total) of the lines between 15-18 months of age. Based on data generated from the current phase of KOMP2, we expect about 30% of lines to be non-viable. We will characterize the non-viable mutants using high-throughput imaging modalities at three embryonic time points. All data generated from embryonic, juvenile and adult mice will be rapidly deposited into the Data Coordination Center (DCC) that supports KOMP2 and the IMPC. Lastly, JAX will work collaboratively with the KOMP2 Regional Network and with member organizations of the IMPC to share protocols, innovation, and new technology and to broadly and openly disseminate our findings to the international community through publication, presentations at meetings, web activities and social media.