Periodic Reporting for period 2 - DNA-Robotics (DNA-Based Modular Nanorobotics) Reporting period: 2020-01-01 to 2022-06-30 Summary of the context and overall objectives of the project Robots are everywhere. They mow your lawn, vacuum your floor and make your new car, just to mention a few examples. No matter their function, they all consist of sensors and actuators and they are currently transforming the way we work and live.The DNA-Robotics consortium have aimed to bring robots into the nanometer scale and have taken the first steps towards the use of molecular robots in future medical and technological applications. In DNA-Robotics we have taken advantage of DNA’s sequence programmability to design self-assembling structural and functional modules that have been combined to perform certain functions. In essence, DNA can be designed and used for building into any arbitrary shape and structure at the nanoscale, and the designed structures can in the future be combined and integrated into even more complex systems. With this in mind, leading European research groups in the field of DNA nanotechnology collaborated to exploit the advantages of DNA programmability and have taken the field to the next level by developing the field of DNA robotics. Each of the research groups had expert knowledge within different topics ranging from physics over chemistry to molecular biology and this combination have created a synergy effect, which have both increased the impact and interdisciplinarity of the project.To achieve the goal of creating modules for a DNA-based robot, we divided the project into different work packages, as describe below:- Integrative structural design- Programming and signal transfer- Sensing and signalling- Actuation- TranslocationIn addition to the design of different modules for the above functions, we combined the modules and explored the integrated function in a biological setting.An integral and very important part of the DNA-Robotics project have been the education of 15 early stage researchers (ESRs). For a period of 36 months, the ESRs have been working at seven different locations with one of the nine principal investigators as their mentor and supervisor. During their employment, the network have ensured that they been offered high-quality training, which have included network training events, secondments within the network, and participation in scientific conferences in addition to the predefined hands-on training project at their home institution. Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far The 15 early stage researchers (ESRs), have been working on the predefined projects and along with their scientific maturation they have actively taken ownership of their individual projects. The DNA-Robotics as a whole has been affected by the COVID-19 pandemic and especially the network meeting and travels have been severely limited. Nevertheless, we were able to go through with all planed network events with 2 workshops as hybrid formats and 2 workshops in a purely virtual format.Each of the 15 ESRs have contributed to our online blog on a regular basis with posts about themselves and their background but also on predefined topics as part of our online lecture series and the online science exhibition. They have further taken part in conferences e.g. FDN2020, FNANO2021 and the final meeting in Aarhus where they presented their work both orally and through a poster session with visitors from the public. Scientifically, the ESRS have all made tremendous progress in their research projects in accordance to the work packages. Which have been clearly visible from the scientific papers that have been published from the different groups. A few examples of our scientific achievements include:- A mechanistically functional DNA origami Bowden cable for actuation- Design and simulation tool for DNA, RNA and hybrid protein–nucleic acid nanostructures - Functional translocation module based on rotary protein- Development of a DNA molecular printer capable of programmable positioning and patterning in two dimensions- DNA origami plasmonic sensingDuring the time of the DNA-Robotics project we have bolstered the awareness of the research area both in the scientific community, industry and to the broader public. A number of high quality papers have already been produced by the ESRs, generating more than 20.000 views, more than a 110 tweets on the papers and the several media outlets have further picked up on some of the science in the papers. The data generated have also been presented at several international conferences e.g. the FDN2020 and FNANO 2021. Finally, our DNA-Robotics homepage have had more than 11.000 visitors during the project period and more have directly been informed about the work on social media e.g. via twitter where the project and the individual groups have had more than 4000 followers combined. So far, a patent including work inspired from DNA-Robotics have been filed and an associated company ATDBio have been acquired by another company in deal worth 45 million GBP.It is our strong expectation that the dissemination and exploitation of the work made in DNA-Robotics will continue to generate considerable impact in the coming years and well beyond the project period. Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far) In DNA-Robotics, we had an ambitious aim to create the modules for a plug-and-play platform, beyond the state of the art, that would allow an end user to go in to a "catalogue" of DNA structures and choose the specific modules necessary for a given purpose. In the project we have created new designs and mechanisms for construction of DNA-based nanostructures and have made significant progress beyond the state of the art on the development of new communication pathways, translocation systems and sensing systems.On the scientific level much of the work has been published in peer-review journals with more manuscripts in review and in preparation. The published work will help to advance new technological breakthroughs in particular in Nano-Robotics but also in Nanoscience and Nanomedicine in general. Knowledge and patents generated from the work have contributed in established companies and may further contribute to the creation of jobs in future start-up companies. We have finally through DNA-Robotics been able to train 15 high level researchers that may continue to contribute to the scientific and broader society with their human capital and acquired technical skills.