High-throughput RNA isoform sequencing using programmed cDNA concatenation.

Al'Khafaji, Aziz M; Smith, Jonathan T; Garimella, Kiran V; Babadi, Mehrtash; Popic, Victoria; Sade-Feldman, Moshe; Gatzen, Michael; Sarkizova, Siranush et al. · Nat Biotechnol · 2024

basic_science · Level V

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Abstract

Full-length RNA-sequencing methods using long-read technologies can capture complete transcript isoforms, but their throughput is limited. We introduce multiplexed arrays isoform sequencing (MAS-ISO-seq), a technique for programmably concatenating complementary DNAs (cDNAs) into molecules optimal for long-read sequencing, increasing the throughput >15-fold to nearly 40 million cDNA reads per run on the Sequel IIe sequencer. When applied to single-cell RNA sequencing of tumor-infiltrating T cells, MAS-ISO-seq demonstrated a 12- to 32-fold increase in the discovery of differentially spliced genes.

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