A high-throughput and quantitative method to assess the mutagenic potential of translesion DNA synthesis

David J. Taggart, Terry L. Camerlengo, Jason K. Harrison, Shanen M. Sherrer, Ajay K. Kshetry, John Stephen Taylor, Kun Huang, Zucai Suo

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

Cellular genomes are constantly damaged by endogenous and exogenous agents that covalently and structurally modify DNA to produce DNA lesions. Although most lesions are mended by various DNA repair pathways in vivo, a significant number of damage sites persist during genomic replication. Our understanding of the mutagenic outcomes derived from these unrepaired DNA lesions has been hindered by the low throughput of existing sequencing methods. Therefore, we have developed a cost-effective high-throughput short oligonucleotide sequencing assay that uses next-generation DNA sequencing technology for the assessment of the mutagenic profiles of translesion DNA synthesis catalyzed by any error-prone DNA polymerase. The vast amount of sequencing data produced were aligned and quantified by using our novel software. As an example, the high-throughput short oligonucleotide sequencing assay was used to analyze the types and frequencies of mutations upstream, downstream and at a site-specifically placed cis-syn thymidine-thymidine dimer generated individually by three lesion-bypass human Y-family DNA polymerases.

Original languageEnglish
Pages (from-to)e96
JournalNucleic acids research
Volume41
Issue number8
DOIs
StatePublished - Apr 2013

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