PubReading [197] - Nanopore microscope identifies RNA isoforms with structural colours - F. Bošković and U. Keyser

<p>Identifying <strong>RNA transcript isoforms</strong> requires intricate protocols that suffer from various enzymatic biases. Here we design three-dimensional molecular constructs that enable identification of transcript isoforms at the <strong>single-molecule level</strong> using solid-state nanopore microscopy. We refold target RNA into RNA identifiers with designed sets of complementary DNA strands. Each reshaped molecule carries a unique sequence of structural (pseudo)colours. Structural colours consist of DNA structures, protein labels, native RNA structures or a combination of all three. The sequence of structural colours of RNA identifiers enables simultaneous identification and relative quantification of multiple RNA targets without prior amplification. Our Amplification-free RNA TargEt Multiplex Isoform Sensing (<strong>ARTEMIS</strong>) method reveals structural arrangements in native transcripts in agreement with published variants. ARTEMIS discriminates circular and linear transcript isoforms in a one-step, enzyme-free reaction in a complex human transcriptome using single-molecule read-out.</p><p><em><a href="https://doi.org/10.1038/s41557-022-01037-5">https://doi.org/10.1038/s41557-022-01037-5</a> -2022</em></p>