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Determining the Location of DNA Modification and Mutation Caused by UVB Light in Skin Cancer

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Technical Report,15 Aug 2012,14 Aug 2015

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University of Colorado Denver Aurora United States

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Extrinsic modifications to the DNA bases such as pyrimidine dimers can arise from a variety of exposures and can lead to aberrant cell growth or death. A detailed view of this base modification is necessary for a more complete view of genetic and epigenetic regulation but the process is poorly understood. We have developed a method to look at the precise genomic position of these modifications using a next generation sequencing approach. During this research period we have optimized this protocol to obtain libraries from UVB damaged human cells. We have purified or obtained the enzymes needed for aim 1 of this research. Using these enzymes we have shown that we obtain cleavage patterns consistent with the administered UV dosage and that sequencing libraries generated for both yeast and human cells show pyrimidine bias on the 5 end, indicating that we are sequencing the dimers. Understanding where these modifications occur is a critical first step to understanding the mutations they cause.

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