How do molluscs build their shells? Deciphering calcium transport mechanisms in shellfish biomineralisation using genome-editing

海洋无脊椎动物已经回答了生物学中最基本的问题。水母提供了第一个荧光蛋白，鱿鱼的巨大轴突告诉我们神经信号是电的，海胆解开了控制早期动物发育的基因调控网络。生物研究受益于多样性，然而，目前主要的模型只包括三种物种：苍蝇、老鼠和斑马鱼。该项目旨在在一种新型贝类模型中开发CRISPR-Cas9介导的基因组编辑方案，以回答生物矿化中钙转运的基本问题。最近的研究成功地证明了CRISPR-Cas9介导的三种软体动物的基因组编辑[1-3]。以这些方案为起点，您将针对候选钙转运基因进行系统敲除，以解决贝类生物矿物生产中有争议的争论：钙是通过细胞（跨细胞）还是在细胞之间（细胞旁）转运到壳中？该奖学金的目标是在工业界和学术界都非常需要的方法开发技能。您将开发的工具- CRISPR-Cas9介导的贝类基因组编辑-是水产养殖和生物技术的理想工具，因为它将促进生产具有增强理想特性（更快，更有效的生长，抗病能力，更坚固的外壳）的转基因动物。在美国，AquaBounty的人类消费转基因鲑鱼可以在16个月内达到市场规模（相比之下，野生型需要3年）。来自非水产养殖模式物种（斑马鱼）的大量研究和技术促进了转基因鲑鱼的生产，为了将这项技术应用于水产养殖贝类物种，必须为软体动物开发一种基因激活的模式系统。你们要解决的研究问题集中在钙的运输上。使用CRISPR-Cas9，您将系统地破坏跨细胞和细胞旁钙运输以及生长培养基中的可用钙，以测试软体动物钙化中使用的起源和运输机制。您将解决钙是通过细胞（跨细胞）运输到外壳，还是在细胞之间（细胞旁）运输，或者，如果钙没有主动的有机运输，而钙在海水中浓度足够高，可以被动沉淀。由于与水产养殖和生物技术的相关性（通过工具开发），以及更广泛的领域，如入侵物种修复（Crepidula fornicata在英国是侵入性和破坏性的，CRISPR-Cas9可用于开发基因驱动修复技术），有空间将您的研究引导到更多的应用途径。为了帮助探索您的工具和知识的更广泛应用，您将与工业CASE合作伙伴- Mikota Ltd.合作。你将加入阿伯丁大学的Sleight实验室（http://sleightlab.com/），这是一个鼓励和培养好奇心的支持性环境。您将接受生物信息学、分子生物学、动物培养和胚胎学（包括显微注射）方面的严格培训。此外，您还可以申请国际暑期学校进行高级培训（如世界著名的美国海洋生物实验室胚胎学课程），并参加国际会议，展示您的研究成果。您将在领先的研究期刊上发表您的研究成果，并在毕业时拥有适合学术界，工业界或更广泛领域的职业生涯的记录。

Marine invertebrates have answered the most fundamental questions in biology. Jellyfish provided the first fluorescent protein, the giant axons of squid taught us nerve signalling is electrical and sea urchins untangled gene regulatory networks controlling early animal development. Biological research benefits from diversity however, current major models include just three species: fly, mouse and zebrafish. This project seeks to develop CRISPR-Cas9 mediated genome-editing protocols in a novel shellfish model to answer a fundamental question on calcium transport in biomineralisation.Recent studies have successfully demonstrated CRISPR-Cas9 mediated genome-editing in three mollusc species [1-3]. Using these protocols as a starting-point, you will target candidate calcium transport genes for systematic knock-out in order to resolve a controversial debate in shellfish biomineral production: is calcium transported to the shell through cells (transcellular) or, in-between cells (paracellular)?This studentship is targeted at a method development skillset that is highly desirable in industry and academia alike. The tool you will develop - CRISPR-Cas9 mediated genome-editing in shellfish - is desirable to aquaculture and biotechnology as it will facilitate the production of genetically modified (GM) animals with enhanced desirable traits (faster more efficient growth, disease resistance, stronger shells). In the USA AquaBounty's human consumption approved transgenic salmon can reach market size in 16 months (versus 3 years for wild-type). The production of GM salmon was facilitated by a wealth of research and technologies from non-aquaculture model species (zebrafish) and, to bring this technology to aquaculture shellfish species, a genetically-enabled model system must be developed for molluscs.The research question you will tackle focusses on calcium transport. Using CRISPR-Cas9 you will systematically disrupt transcellular and paracellular calcium transport in addition to available calcium in the growth medium to test the origin and transport mechanism used in molluscan calcification. You will resolve weather calcium is transported to the shell through cells (transcellular), in-between cells (paracellular) or, if there is not active organismal transport of calcium, and instead calcium is available in high enough concentrations in seawater to precipitate passively.Owing to the relevance to aquaculture and biotechnology (via the tool development), as well as wider fields such as invasive species remediation (Crepidula fornicata is invasive and damaging in the UK, CRISPR-Cas9 could be used to develop gene-drive remediation technology) there is scope to direct your research to more applied avenues. In order to aid exploration into the wider applications of your tools and knowledge, you will work with an industrial CASE partner - Mikota Ltd. You will join the Sleight Lab (http://sleightlab.com/) at the University of Aberdeen, a supportive environment where curiosity is encouraged and nurtured. You will receive rigorous training in bioinformatics, molecular biology, animal culture and embryology (including microinjection). In addition, you will be supported to apply for international summer schools for advanced training (such as the world-famous Embryology Course at the Marine Biology Laboratory USA), and attend international conferences to present your research. You will publish your findings in leading research journals and graduate with a track-record apt for a career in academia, industry or wider fields.