DFG 460736965 - Array Synthesizer

Biological photolithography uses optical approaches from the integrated semiconductor industry in combination with slightly modified solid-phase synthesis chemistry to enable the chemical synthesis of biological oligomers on an ultra-large scale and their use for multiplexing bioaffinity assays or for the generation of extremely large sequence-determined libraries. Oligopeptide synthesis on this scale enables the efficient development of epitope-specific antibodies and the characterization of protein functions and protein-protein interactions relevant to cellular regulation and signaling. Applications of large-scale nucleic acid synthesis include gene expression analysis, gene assembly, DNA origami, spatial transcriptomics, and digital information storage. Due to the complexity and proprietary nature of most biotechnological processes, large-scale synthesis approaches are currently limited to commercial services. The aim of this project is to develop an open-source benchtop device that enables the parallel synthesis of millions of unique sequences. This device will be a next-generation maskless array synthesizer, an optical device coupled to a standard solid-phase synthesizer that provides robust, simple, and low-cost access to ultra-large-scale synthesis, particularly of nucleic acids and peptides, but also facilitates the development of synthesis approaches suitable for other oligomers, such as oligosaccharides or engineered biomimetic constructs. To demonstrate the performance and versatility of the highly efficient new device, we will (1) apply ultra-large-scale photolithographic nucleic acid synthesis to generate DNA libraries for digital data archiving. In particular, we will show that ultra-large-scale synthesis combined with efficient encoding leads to cost-effective and efficient molecular-level information storage and retrieval from DNA. Furthermore, in combination with our recently developed photolithographic approach to RNA synthesis, we will (2) synthesize extremely large permutative RNA libraries to comprehensively explore the binding affinity and fluorescence-enhancing sequence landscape of the mango family of fluorogenic RNA aptamers.

DFG

New equipment for research