An x-ray irradiator cabinet for live cell irradiation

每个活着的有机体的细胞都不断地暴露在DNA损伤中，无论是通过内源性新陈代谢的副产品产生的，还是由辐射或化学物质等细胞外来源产生的。虽然DNA损伤是常见的，但它可能导致突变，对细胞、组织和整个生物体产生负面影响。不能修复DNA损伤会导致多种疾病，包括免疫缺陷、衰老、癌症和神经退行性疾病。因此，了解细胞如何对DNA损伤做出反应是理解细胞和物种长期和短期生存所必需的基本机制的关键。 这笔赠款提供的资金将允许购买一台Faxitron CellRad柜式X射线装置，该装置可为细胞和组织培养物照射提供一系列辐射剂量。细胞辐射会对细胞的遗传物质(DNA)造成损害。这是一个实验模型系统，用于研究哺乳动物和人类细胞修复其受损DNA的机制。这一设备对于开展斯柴尔德-保尔特实验室由NSERC资助的研究项目至关重要，该项目调查DNA修复蛋白Ku的作用，该蛋白对修复哺乳动物细胞中的DNA双链断裂至关重要。虽然人们知道Ku对于DNA的正确修复是必不可少的，但它的作用模式仍然不清楚。众所周知，Ku与许多其他因素相互作用，这些因素在DNA修复中发挥作用，然而，这些相互作用的细节和结果尚不清楚。利用Ku的晶体结构，人们可以预测蛋白质中可以接触到其他蛋白质的区域。因此，在这些可访问的区域中的几个产生了突变，以测试它们如何改变Ku DNA修复功能。已经确定了两个Ku突变，它们改变了其对DNA损伤的功能。这些突变导致不同的缺陷，表明突变区域涉及不同的功能。斯柴尔德-保尔特实验室目前正在努力鉴定与这些Ku基序相互作用的蛋白质，以阐明它们如何在DNA修复中共同发挥作用。 所要求的设备将提供在我们的细胞模型中诱导DNA损伤的手段，以便我们能够阐明Ku在DNA修复中的调节机制。 西部大学的其他几个实验室也需要这种设备来研究干细胞再生和细胞介导的免疫反应。因此，这种设备造成的DNA损伤是一种工具，使学生、科学家和技术人员在各自的实验室或研究项目中进行生物研究时使用的生物系统和实验。

The cells of every living organism are constantly being exposed to DNA damage, whether created through by-products of the endogenous metabolism, or generated by extracellular sources such as radiation or chemicals. While DNA damage is commonly encountered, it can cause mutations that have negative impacts on cells, tissues, and organisms as a whole. Failure to repair DNA lesions results in a multitude of conditions including immunodeficiency, aging, cancer, and neurodegenerative disorders. Thus, understanding how cells respond to DNA damage is key to comprehend fundamental mechanisms that are essential to long- and short-term survival of cells and species. The funding provided by this grant will allow the purchase a Faxitron CellRad cabinet X-ray unit that delivers a range of radiation doses for cell and tissue culture irradiation. Cell irradiation causes damage in the genetic material (DNA) of the cells. This serves as an experimental model system to study the mechanisms set in place by mammalian and human cells to repair their damaged DNA. This equipment is essential to carry out the Schild-Poulter lab’s research program funded by NSERC that investigates the role of the DNA repair protein Ku which is essential for the repair of DNA double-stranded breaks in mammalian cells. While it is understood that Ku is essential for the proper repair of DNA, its mode of action is still unclear. It is known that Ku interacts with a number of other factors that play roles in the repair of DNA, however, the details and the outcome of these interactions are poorly understood. Using the crystal structure of Ku, one can predict regions of the protein that are accessible to contact other proteins. Thus, mutations were generated in several of these accessible regions to test how they alter Ku DNA repair functions. Two Ku mutations that alter its ability to function in response to DNA damage have been identified. These mutations result in different defects suggesting that the mutated regions are involved in distinct functions. The Schild-Poulter laboratory is currently working to identify the proteins that interact with these Ku motifs to elucidate how they function together in the repair of DNA. The requested equipment will provide the means to induce DNA damage in our cellular models so that we can elucidate the regulatory mechanisms that underlie Ku function in DNA repair. This equipment is also needed by several other laboratories at Western University for research on stem cell regeneration and cell-mediated immune responses. Thus, the DNA damage created by this equipment is a tool to enable biological systems and experimentations used in biological research by students, scientists and technicians in their individual lab or research program.