Integrated System for Editing Eukaryotic Genomes

使用工程“巨核酸酶”，特别是基于CRISPR/Cas系统的巨核酸酶的最新发展，使需要有针对性的基因修改的研究发生了革命性的变化。这项技术极大地提高了对细胞和动物进行基因工程所需的效率、成本和时间。我们实验室(Bordignon和Agellon)的研究已经证实CRISPR/CAS系统在细胞和发育中的胚胎中都是有效的，我们目前正在将这项技术应用于特定于细胞/组织的应用。为了充分利用这项技术，需要一种综合的方法和系统来简化从基因编辑到基因组编辑动物生产的过程。我们的团队由麦吉尔大学两个院系的三个系在各自领域内公认的知名领导者组成。总的来说，我们拥有多学科的专业知识，从分子到整个动物生理学，并有共同的研究兴趣和目标。共同申请者之间正在进行的研究合作包括：细胞重新编程和发育(Bordignon，Agellon，Duggavathi，Clarke)；细胞应激和应对机制(Agellon，Bordignon)；代谢和营养(Agellon，赵)；动物生育和代谢(Duggavathi，Bordignon，Agellon)；肠道病理生理学、宿主/病原体相互作用和组织反应(赵，Bordignon，Agellon)；卵泡发生、卵子发生和胚胎发育的遗传和表观遗传调控(Clarke，Bordignon，Duggavathi)。我们实验室的研究得到了私人“行业”和公共来源的支持，包括NSERC(Discovery、CRDPJ、Engage)、CIHR、FRQNT和CFI。我们要求支持校园内的基因组编辑集成系统，以支持基础研究和翻译研究，涉及基因编辑模型的生成和使用，包括细胞、胚胎和活动物。这一建议的系统包括允许操纵单个细胞和胚胎的独特组件，向细胞器、细胞和胚胎中微量注射毫微升体积，在精确培养条件下支持操纵的细胞和胚胎。拟议的系统将使我们能够对不同物种的培养细胞和胚胎进行基因组编辑，以研究发育、新陈代谢和病理生理过程，以及创造用于基础、生物医学和翻译研究的基因编辑动物。使用这种最先进的集成系统将提高能力，并使新的研究合作成为可能，从而为麦吉尔公司内部以及国内和国际学术界、政府和行业的HQP提供一个高度丰富的多学科培训环境。

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.