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Automated high-throughput selection of DNA aptamers using a common optical next-generation sequencer

Drees, A.; Ahlers, C.; Kehrer, T. P.; Ehmke, N.; Gruen, A. F.-R.; Uetrecht, C.; Ignatova, Z.; Schumacher, U.; Fischer, M.

2024-06-25 biochemistry
10.1101/2024.06.24.600375 bioRxiv
Show abstract

Aptamers are conventionally selected via Systematic Evolution of Ligands by Exponential Enrichment (SELEX). This process is, however, laborious, time-consuming, and has a relatively low efficacy. Here, we developed an automated high-throughput screening platform for the selection of DNA aptamers which consists of an optical next-generation sequencer with a modified software and hardware to automatically perform fluorescence-based binding assays on the displayed DNA sequences subsequent to sequencing. Using this platform, after only three to five SELEX rounds we selected highly affine DNA aptamers for the lectins LecA and LecB of Pseudomonas aeruginosa as well as for the Pseudomonas Exotoxin A. In comparison, twelve rounds of conventional SELEX resulted in three-fold less affine aptamers for LecA and PEA and none for LecB. Our high throughput-approach bears great potential to augment SELEX as it significantly increases time efficiency, enabling the selection of aptamers within only one week.

Published in Nucleic Acids Research (predicted rank #2) · training set

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