Language

English

Publication Date

10-3-2024

Journal

Molecular Cell

DOI

10.1016/j.molcel.2024.07.008

PMID

39096899

PMCID

PMC11455606

PubMedCentral® Posted Date

10-3-2025

PubMedCentral® Full Text Version

Author MSS

Abstract

Despite the numerous sequencing methods available, the diversity in RNA size and chemical modification makes it difficult to capture all RNAs in a cell. We developed a method that combines quasi-random priming with template switching to construct sequencing libraries from RNA molecules of any length and with any type of 3' modifications, allowing for the sequencing of virtually all RNA species. Our ligation-independent detection of all types of RNA (LIDAR) is a simple, effective tool to identify and quantify all classes of coding and non-coding RNAs. With LIDAR, we comprehensively characterized the transcriptomes of mouse embryonic stem cells, neural progenitor cells, mouse tissues, and sperm. LIDAR detected a much larger variety of tRNA-derived RNAs (tDRs) compared with traditional ligation-dependent sequencing methods and uncovered tDRs with blocked 3' ends that had previously escaped detection. Therefore, LIDAR can capture all RNAs in a sample and uncover RNA species with potential regulatory functions.

Keywords

Animals, RNA, Transfer, Mice, Sequence Analysis, RNA, Male, Mouse Embryonic Stem Cells, Transcriptome, Neural Stem Cells, High-Throughput Nucleotide Sequencing, Spermatozoa

Published Open-Access

yes

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