Ribosome profiling to reveal how temperature affects protein synthesis in diatoms

核糖体分析揭示温度如何影响硅藻中的蛋白质合成

• 批准号: 2116995
• 金额: --
• 依托单位: University of East Anglia
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2116995
• 关键词: Ribosome; profiling; reveal; how; temperature

Diatoms are important microalgae with high biotechnological potential. Several aspects of diatom physiology including the silica frustule, lipid storage and photosynthesis are being applied to biotechnology. Areas of interest include nanotechnology, drug delivery, biofuels, solar capture and bioactive compounds. Given the ecological importance of diatoms and their applications for biotechnology, several different genetic tools have recently been developed in the Mock lab for the model diatom Thalassiosira pseudonana such as genome editing by CRISPR/Cas9 and ribosome profiling. The application of both tools in this project will enable the PhD student to obtain fundamental insights into how temperature affects translation in the model diatom Thalassiosira pseudonana. Since there are no data available yet on how temperature regulates protein synthesis in any algae on a mechanistic level, we suggest to apply ribosome profiling to provide fundamental insights into if and how temperature affects a) the location of translation start sites, b) the density of ribosomes on messenger RNAs and c) the speed of translating ribosomes. Furthermore, the role of codon usage and its impact on tRNA evolution in relation to the recently discovered tRNA-derived small non-coding RNAs for protein synthesis in diatoms will be investigated using the genome editing tool CRISPR/Cas in combination with ribosome profiling. We aim to modify the genetic code in T. pseudonana in order to obtain first insights into codon usage, tRNA expression and the role of tRNA-derived non-coding RNAs. Data from this project will lay the foundation for synthetic biology with diatoms as translation underpins the synthesis of various different enzymes and materials (e.g. silica) used in algal biotechnology.

硅藻是重要的微藻，具有很高的生物技术潜力。硅藻生理学的几个方面，包括硅藻壳、脂质储存和光合作用，正在应用于生物技术。感兴趣的领域包括纳米技术、药物输送、生物燃料、太阳能捕获和生物活性化合物。鉴于硅藻的生态重要性及其在生物技术中的应用，模拟实验室最近为硅藻模型假微型海链藻开发了几种不同的遗传工具，例如通过 CRISPR/Cas9 进行基因组编辑和核糖体分析。这两种工具在该项目中的应用将使博士生能够获得关于温度如何影响硅藻模型假微型海链的翻译的基本见解。由于目前还没有关于温度如何在机制水平上调节任何藻类中蛋白质合成的数据，我们建议应用核糖体分析来提供关于温度是否以及如何影响 a) 翻译起始位点的位置、b) 信使 RNA 上核糖体的密度和 c) 核糖体翻译速度的基本见解。此外，将使用基因组编辑工具 CRISPR/Cas 结合核糖体分析来研究密码子使用的作用及其对最近发现的用于硅藻蛋白质合成的 tRNA 衍生小非编码 RNA 的 tRNA 进化的影响。我们的目标是修改 T.pseudonana 的遗传密码，以便初步了解密码子使用、tRNA 表达和 tRNA 衍生的非编码 RNA 的作用。该项目的数据将为硅藻合成生物学奠定基础，因为翻译支撑着藻类生物技术中使用的各种不同酶和材料（例如二氧化硅）的合成。