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

项目摘要 设备补充申请要求资金购买Bio-Rad实时PCR仪 这对于OGG 1基因敲除和胃L3基因敲除的验证以及 绘制m6 A和RNA的分布图，以及DNA聚合酶β（Pol β）基因的DNA损伤， 根据PI的NIEHS主动资助R 03 ES 0350200的目标，链断裂介导的 用于m6 A和RNA作图的RNA和DNA修饰分析测定（SBRM和SBRM）， Pol β基因上的DNA损伤是基于PCR的，并且对于所提议的研究的成功至关重要。 PI实验室的旧实时PCR仪最近坏了。无法修复 因为旧型号及其维修服务已被Bio-Rad实验室停止。一个新的真实的- 时间PCR机将使我们能够实现拟议的研究目标，了解 RNA和DNA损伤对m6 A沉积在Pol β基因上的影响以及m6 A谱如何变化 可以通过调节癌细胞中Pol β转录物上的RNA损伤来改变DNA修复。因此， 迫切需要购买新的实时PCR仪，这对我们完成 我们的研究目标按时完成。

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.