Lab robotics for synthetic biology and strain evaluation (RoboSyn)

实验室机器人RoboSyn应该用于斯图加特大学的合成生物学和系统生物学研究，特别是在生物研究所(IBVT)。该设备应支持(A)菌株和过程工程、(B)无细胞蛋白质合成和(C)RNA-RNA相互作用的关键研究活动。通过建立RoboSyn，将弥合目前在实验室自动化以及在毫升和米克罗L规模的并行菌株测试方面的差距。RoboSyn的设计使体外和体内测试都能成功进行。这项技术建立在申请者的专业知识和在斯图加特建立的技术基础上。RoboSyn通过提供实验室自动化来补充现有的基础设施，这是到目前为止还没有的。此外，从DNA片段中产生编码DNA并将其整合到微生物宿主中的现代方法将实现自动化。研究还将包括通过细胞裂解试验优化单个基因的表达水平(例如，在生物合成途径中)。这个问题与(B)的主题有关，即细胞自由蛋白质合成。基于深厚的前期研究专业知识，新的细胞和表达宿主将成为目标。详细的系统生物学研究将阐明它们的功能以及它们是否适合于生产具有药物和医学吸引力的化合物。此外，RoboSyn将用于阐明原核细胞和真核细胞(C)中的RNA-RNA介导的调控网络。这类系统生物学研究将受益于在并行读出系统中安装不同的扰动并随后进行“全局”监测的可能性。与IBVT的主旨一致，所有研究都将利用数学模型来检查实验数据或设计实验。因此，研究将被构建为完整的设计-建造-测试-学习周期。后者特别重要，因为它确保对菌株和过程进行基于知识的优化。

The lab robotic RoboSyn should be used for synthetic biology and systems biology research at the University of Stuttgart, in particular at the ‘Institut für Bioverfahrnstechnik (IBVT). The device should support the key research activities (A) strain- and process engineering, (B) cell-free protein synthesis, and (C) RNA-RNA interaction. By establishing RoboSyn the current gap in lab automation and in parallel strain testing in mL and mikroL scale will be closed. RoboSyn is designed such that in vitro and in vivo tests can be performed successfully. The technology builds on the expertise of the applicants and on the knowhow established in Stuttgart. RoboSyn complements existing infrastructure by giving access to lab automation which is missing so far.Regarding (A) RoboSyn will be used for the massively parallel evaluation of novel strains constructed in multiple projects. Besides, modern approaches of generating coding DNA from DNA fragments and their integration in microbial hosts will be automatized. Studies will comprise the optimization of expression levels of individual genes (e.g. in biosynthetic pathways) via cell lysate tests, too. The question is linked to the topics of (B), the cell free protein synthesis. Based on profound expertise of preliminary studies, novel cells and expression hosts will be targeted. Detailed systems biology studies will elucidate their functioning and their suitability for the production of pharmaceutically and medically attractive compounds. Furthermore, RoboSyn will be used to elucidate RNA-RNA mediated regulatory networks in pro- and eukaryotic cells (C). Such systems biology studies will benefit from the possibility of installing distinct perturbations followed by ‘global’ monitoring in the parallel read-out system.In agreement with the leitmotif of IBVT, all studies will make use of mathematical models either to check experimental data or to design experiments. As such, studies will be structured as complete design-build-test-learn cycles. The latter is of particular importance as it ensures a knowledge-based optimization of strains and processes.