High-throughput RNA Isoform Sequencing Using Programmed CDNA Concatenation

Overview

Journal Nat Biotechnol

Specialty Biotechnology

Date 2023 Jun 8

PMID 37291427

Authors

Aziz M AlKhafaji

Jonathan T Smith

Kiran V Garimella

Mehrtash Babadi

Victoria Popic

Moshe Sade-Feldman

Michael Gatzen

Siranush Sarkizova

Marc A Schwartz

Emily M Blaum

Allyson Day

Maura Costello

Tera Bowers

Stacey Gabriel

Eric Banks

Anthony A Philippakis

Genevieve M Boland

Paul C Blainey

Nir Hacohen

Affiliations

Soon will be listed here.

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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