Duncan NRI Faculty and Staff Publications

Language

English

Publication Date

5-1-2024

Journal

Nature Microbiology

DOI

10.1038/s41564-024-01655-4

PMID

38649410

PMCID

PMC11384275

PubMedCentral® Posted Date

5-1-2025

PubMedCentral® Full Text Version

Author MSS

Abstract

RNA viruses, like SARS-CoV-2, depend on their RNA-dependent RNA polymerases (RdRp) for replication, which is error-prone. Monitoring replication errors is crucial for understanding the virus’s evolution. Current methods lack the precision to detect rare de novo RNA mutations, particularly in low-input samples such as those from patients. Here, we introduce a new targeted Accurate RNA Consensus sequencing method (tARC-seq) to accurately determine the mutation frequency and types in SARS-CoV-2, both in cell culture and clinical samples. Our findings show an average of 2.68×10−5 new errors per cycle with a C>T bias that cannot be solely attributed to APOBEC editing. We identified hotspots and cold spots throughout the genome, correlating with high or low GC content, and pinpointed transcription regulatory sites as regions more susceptible to errors. tARC-seq captured template switching events including insertions, deletions, and complex mutations. These insights shed light on the genetic diversity generation and evolutionary dynamics of SARS-CoV-2.

Keywords

SARS-CoV-2, Humans, Virus Replication, COVID-19, Genome, Viral, RNA, Viral, Mutation, Sequence Analysis, RNA, Evolution, Molecular, Mutation Rate

Published Open-Access

yes

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