Infection free and scalable production of DNA oligonucleotides for mass production of nano-objects by self-assembling

生物体的研究是DNA分子广泛应用于纳米技术的下一个里程碑。对于后者以及DNA纳米物体在材料科学中的应用研究，单链DNA（ss-DNA）的可扩展供应将需要千克级。因此，需要新的ss-DNA生产工艺，以使从克到千克规模甚至更远的一步。为了能够有效地生物技术大规模生产确定的ss-DNA对象，必须利用现有的生物技术生产工厂。合同制造商不可能使用感染性噬菌体M13生产ss-DNA支架，或使用辅助噬菌体与噬菌粒组合生产用户定义的人工ss-DNA，因为在所有其他生产过程中，不需要的噬菌体感染风险持续存在。杆菌因此，本项目的目的是合理设计一种新的方法，用于在单锅反应中无感染和有效地生产用户定义的ss-DNA，用于纳米物体的自组装。因此，在大肠杆菌中，应该使用辅助质粒和噬菌粒来代替感染性辅助噬菌体。杆菌

Studies in living organisms are the next milestone on the way to broad applicability of DNA-molecules for nanotechnology. For the latter as well as for studies on the application of DNA nano-objects in material sciences, the scalable supply of single-stranded-DNA (ss-DNA) will be necessary on a kilogram scale. Thus new ss-DNA production processes are required to make the step forward from gram to kilogram scale and even further. To enable the efficient biotechnological mass production of defined ss-DNA objects, it is essential to make use of existing biotechnological production plants. Using the infectious bacteriophage M13 for ss-DNA scaffold production or using a helperphage in combination with a phagemid for the production of user-defined and artificial ss-DNA with Escherichia coli is impossible with contract manufacturers, as the risk of unwanted phage infections persists throughout all other production processes performed there with E. coli. Thus the aim of this project is the rational design of a new process for infection free and efficient production of user-defined ss-DNA for self-assembly of nano-objects in a single-pot reaction. For this, instead of an infectious helperphage, a helperplasmid in combination with a phagemid is supposed to be used in E. coli.