Integrated System for Editing Eukaryotic Genomes

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Recent developments using engineered “meganucleases”, particularly those based on the CRISPR/Cas system, have revolutionized research requiring targeted gene modifications. This technology has dramatically increased the efficiency and reduced the cost and time it takes to genetically engineer cells and animals. Research in our laboratories (Bordignon and Agellon) has confirmed the efficiency of the CRISPR/Cas system in both cells and developing embryos, and we are currently progressing towards adapting this technology for cell/tissue-specific applications. In order to take full advantage of this technology, an integrated approach and system are necessary to streamline the process from gene editing to production of genome-edited animals. Our team is composed of internationally recognized established leaders in their areas representing three departments in two faculties at McGill University. Collectively, we possess multidisciplinary expertise, spanning molecular to whole animal physiology, and have common research interests and goals. Ongoing research collaborations among the co-applicants include: cell reprogramming and development (Bordignon, Agellon, Duggavathi, Clarke); Cell stress and coping response mechanisms (Agellon, Bordignon); metabolism and nutrition (Agellon, Zhao); animal fertility and metabolism (Duggavathi, Bordignon, Agellon); gut pathophysiology, host/pathogen interaction and tissue responses (Zhao, Bordignon, Agellon); genetic and epigenetic regulation of folliculogenesis, oogenesis and embryo development (Clarke, Bordignon, Duggavathi). Research in our laboratories is supported by both private “industry” and public sources, including NSERC (Discovery, CRDPJ, Engage), CIHR, FRQNT and CFI. We request support for an on-campus integrated system for genome editing to support research on fundamental and translational investigation involving the generation and use of gene edited models, including cells, embryos and live animals. This proposed system comprises of unique components allowing for manipulation of individual cells and embryos, microinjection of femtoliter volumes into organelles, cells and embryos, support of manipulated cells and embryos under precise culture conditions. The proposed system will allow us to carry out genome editing in cultured cells and embryos of different species to study development, metabolism and pathophysiology processes, as well as to create gene-edited animals for fundamental, biomedical and translational research. Access to such a state-of-the-art integrated system will increase capacity and enable new research collaborations leading to a highly enriched, multidisciplinary training environment for HQP within McGill and at the national and international levels in academia, government, and industry.