Novel approaches to identify regulators of Bak1 splicing

此应用程序的目标是无偏见地识别编程内在神经元所必需的调节因子 对细胞凋亡的抵抗。细胞凋亡是一种普遍存在的调节细胞死亡的途径，控制着细胞的周转和 后生动物的组织动态平衡。一个长期存在的问题是，神经元如何抑制细胞凋亡，从而有利于 长寿。以往对神经细胞凋亡调控的研究主要集中在外源性存活的原因和方式上。 线索通过控制细胞死亡来帮助建立和维持神经回路。虽然这些鼓舞人心的研究 描绘了神经元在回路形成时如何竞争生存，重要的问题仍然存在 未解决的问题，即神经元的凋亡敏感性是否由基因决定。在我们的初步研究中，我们 已经发现神经元对凋亡的抵抗是在电路形成之前被内在编程的 促凋亡线粒体蛋白BAK1的耗尽。我们进一步发现，BAK1的表达被控制在 通过健壮的替代剪接机制提高RNA水平。关键的问题是：哪些因素计划 Bak1拼接？确定这些因素将产生重大影响，并使新的调查成为可能 神经细胞死亡在不同的环境中控制。拟议的研究将产生新的工具并开发强大的 以细胞为基础的方法系统地识别Bak1剪接的调控因子。我们将把实验和实验相结合 加速发现的计算方法，否则将是有限的和不那么敏感的。我们的 该团队展示了在研究细胞死亡、神经元存活和RNA方面的成功合作 在大脑中的分子遗传学，并期待拟议的研究将取得丰硕成果。通过揭示新的监管机构 细胞凋亡的机制和相关的遗传因素，我们的发现可能为增强细胞的策略提供参考 生存和应对神经退行性疾病。拟议研究的完成也将为我们研究任何替代外显子的广泛适用的战略提供原则证明。

The objective of this application is to unbiasedly identify regulators essential for programming intrinsic neuronal resistance to apoptosis. Apoptosis is a ubiquitous regulated cell death pathway controlling cell turnover and tissue homeostasis in metazoans. A longstanding issue is how neurons suppress apoptosis in favor of longevity. Previous studies on neuronal regulation of apoptosis have focused on why and how extrinsic survival cues help establish and maintain neural circuits through the control of cell death. While these inspiring studies delineate how neurons compete for survival at the time of circuit formation, important questions remain unresolved, i.e., whether neuronal apoptosis sensitivity is genetically determined. In our preliminary studies, we have found that neuronal resistance to apoptosis is intrinsically programmed before circuit formation by depletion of pro-apoptotic mitochondrial protein BAK1. We further found that BAK1 expression is controlled at the RNA level through robust alternative splicing mechanisms. The critical question is: what factors program Bak1 splicing? Identification of these factors will have significant impacts and enable new investigations of neuronal cell death controls in various settings. The proposed study will generate new tools and develop robust cell-based methods to systematically identify regulators of Bak1 splicing. We will integrate experimental and computational approaches to accelerate discoveries that would otherwise be limited and less sensitive. Our team has demonstrated successful collaborations researching cell death, neuronal survival, and RNA molecular genetics in the brain, and expect the proposed research to be fruitful. By revealing novel regulatory mechanisms of apoptosis and associated genetic factors, our findings may inform strategies for enhancing cell survival and tackling neurodegenerative diseases. Completion of the proposed study will also provide proof-of-principle for our broadly applicable strategy to study any alternative exons.