Ribosome profiling to reveal how temperature affects protein synthesis in diatoms

硅藻是一种重要的微藻，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。考虑到硅藻的生态重要性及其在生物技术中的应用，最近在模拟实验室中为模式硅藻Thalassiosira dumerana开发了几种不同的遗传工具，例如CRISPR/Cas9基因组编辑和核糖体分析。这两种工具在这个项目中的应用将使博士生获得基本的见解，温度如何影响模式硅藻海链藻的翻译。由于目前还没有关于温度如何在机械水平上调节任何藻类蛋白质合成的数据，我们建议应用核糖体分析来提供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）翻译核糖体的速度的基本见解。此外，密码子使用的作用及其对tRNA进化的影响与最近发现的tRNA衍生的小非编码RNA在硅藻中的蛋白质合成将使用基因组编辑工具CRISPR/Cas结合核糖体分析进行研究。我们的目标是修改T.为了获得密码子使用，tRNA表达和tRNA衍生的非编码RNA的作用的第一个见解。该项目的数据将为合成生物学奠定基础，硅藻作为翻译支持藻类生物技术中使用的各种不同酶和材料（如二氧化硅）的合成。

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.