N6-methyladenosine (m6A) Interplays with RNA and DNA Damage to Regulate DNA Repair

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Project Summary The equipment supplement application requests funds to purchase a Bio-Rad real-time PCR machine that is essential for the validation of OGG1 gene knockout and METTL3 gene knockdown and for mapping the profiles of m6A and RNA and DNA damage on the DNA polymerase β (Pol β) gene in cancer cells under the aims of the PI’s NIEHS active grant R03ES0350200. The strand break-mediated RNA and DNA modification profiling assays (SBRM and SBRM) used for mapping m6A and RNA and DNA damage on the Pol β gene are PCR-based and critical for the success of the proposed studies. The old real-time PCR machine in the PI’s laboratory was broken recently. It cannot be repaired because the old model and its repair service were discontinued by Bio-Rad Laboratories. A new real- time PCR machine will allow us to accomplish the goals of the proposed studies on understanding the effects of RNA and DNA damage on m6A deposition on the Pol β gene and how m6A profiling changes can alter DNA repair by modulating RNA damage on the Pol β transcript in cancer cells. Thus, there is an urgent need to purchase the new real-time PCR machine that is essential for us to accomplish the research goals of our project on time.