Work

Coumarin-Based Photolabile Solid Support Facilitates Nonchromatographic Purification of RNA Oligonucleotides

Year: 2025

Type: article

Abstract: We describe the synthesis and application of a coumarin-based photocleavable anchor for solid-phase synthesis and nonchromatographic purification of RNA oligonucleotides. In contrast to standard nitroaryl anchors, which are cleaved with ultraviolet (UV) light, the described construct requires visible light (λ = 456 nm) for photocleavage. This is especially important for RNAs that contain a phosphorothioate backbone, which is not compatible with UV light. We optimized the synthesis and purification process using a model 20-nt poly-U RNA and subsequently applied it to structured and unstructured RNA oligonucleotides that contain both phosphodiester and phosphorothioate backbones. Lastly, we purified 103-nt sgRNA and checked its functional fidelity by CRISPR experiments.

Source: The Journal of Organic Chemistry

DOI: 10.1021/acs.joc.5c01528

Authors: Ian McClain, Hilal Dagci, Bhoomika Pandit, Maksim Royzen

Institution: University at Albany, State University of New York

Language: English

Topic: DNA and Nucleic Acid Chemistry

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: hybrid

Estimated APC paid: $4,500

Funders: National Institutes of Health, National Human Genome Research Institute, National Institute of General Medical Sciences