Gene editing and phenotyping suite

我们正在申请一个基因编辑和表型分析套件的资金，该套件将允许我们修改昆虫、两栖动物和鱼类的基因组，并研究这些修改的生物学后果。该设备对NSERC支持的埃文斯、威尔逊、德沃金实验室的研究项目至关重要，这是普鲁特实验室提出的研究主题，并将提供给校园内的其他小组和合作机构。该套装包括几个组件(吸管拉出器、两个微操作器、两个微微管、两个显微镜，并升级到现有的显微镜、两个孵化器和一个非洲爪蛙循环系统)，这些组件将使我们能够在分子、细胞、器官和整个有机体的分辨率水平上生成和研究工程基因组。该套件将允许我们通过使用最先进的方法移除功能(敲除)和插入功能(敲入)来改变动物基因组，并且相对于其他技术具有无与伦比的效率和专一性。从不同的生物(昆虫、两栖动物、鱼类)中产生和分析突变系的能力将使我们能够以一种目前无法实现的方式显著推进我们的研究计划。它将利用我们在无脊椎动物和脊椎动物生物的动物护理设施、基因组学和生物信息学、分子生物学和成像方面所做的大量投资。在不久的将来(未来3年)，这一基础设施将用于研究发育途径的结构和保护、对环境压力和毒素的生理反应、性状的进化和可塑性以及内分泌干扰。从长远来看，根据我们未来的研究结果，它将被部署到广泛的生物操作和实验中。该设施包括将加强HQP培训的功能(例如，带有摄像头和监视器的显微镜；配对的HQP工作流程)。为了确保我们最大限度地延长这套设备的使用寿命，我们将为所有用户制定财务计划和严格的培训计划，包括与公平、多样性和包容性有关的问题。我们迫切需要这种基础设施，因为我们目前无法执行他们在麦克马斯特的研究计划的基本方面，因为没有合适的设备，或者在间歇性访问的非集中地点被充分利用。我们将部署的基因工程方法将使我们能够为理解基本的生物现象做出重要贡献，这些现象是我们各种研究计划的重点。更广泛地说，基因工程为全球范围内的无数问题(疾病、粮食生产、物种生存)带来了巨大的希望。因此，我们的HQP将接受这种设备的培训，这种设备将打开智力之门和实验方法，并为社会带来广泛和持久的潜在好处。

We are requesting funds for a gene editing and phenotyping suite that will allow us to modify the genomes of insects, amphibians, and fish, and study the biological consequences of these modifications. This equipment is critical to the NSERC-supported research programs in the Evans, Wilson, Dworkin labs, a proposed research theme in the Pruitt lab, and will be available to other groups on campus and collaborating institutions. This suite includes several components (pipette puller, two micromanipulators, two picospritzers, two microscopes, and upgrade to an existing microscope, two incubators, and a recirculating system for Xenopus frogs) that together will enable us to generate and study engineered genomes at molecular, cellular, organ, and whole organism level of resolution. This suite will allow us to alter animal genomes both by removing function (knock-outs) and inserting function (knock-ins) using state-of-the-art methods, and with unparalleled efficiency and specificity relative to other techniques. The ability to generate and analyze mutant lines from diverse organisms (insects, amphibians, fish) will enable us to significantly advance our research programs in a way that is not currently possible. It will leverage the considerable investments we have made in animal care facilities for invertebrate and vertebrate organisms, genomics and bioinformatics, molecular biology, and imaging. In the immediate future (next 3 years), this infrastructure will be used to study the structure and conservation of developmental pathways, the physiological response to environmental stresses and toxins, trait evolvability and plasticity, and endocrine disruption. Over the long term it will be deployed for a broad array of biological manipulations and experimentations, as directed by our future research results. This facility includes features that will enhance HQP training (e.g., a microscope with camera and monitor; paired HQP workflow). To ensure we maximize the functional lifespan of this suite of equipment, we will establish both a financial plan and a rigorous training program for all users, including issues pertaining to equity, diversity, and inclusion. This infrastructure is urgently needed because we are currently unable to perform fundamental aspects of their research program at McMaster because suitable equipment is either not available or being fully utilized in a non-centralized locations with intermittent access. The genetic engineering approaches we will deploy will allow us to make important contributions to understanding of fundamental biological phenomena that are the foci of our diverse research programs. More broadly, genetic engineering holds great promise for a myriad of problems with a global scope (disease, food production, species survival). Our HQP thus will receive training with this equipment that will push open intellectual doors and experimental approaches, and with extensive and long lasting potential benefits to society.