A chemical biology toolbox for RNA post-transcriptional modification and capture

摘要 RNA是携带遗传信息和翻译功能蛋白的关键中间体，以及 最近还发现了许多类型的非编码RNA，它们在细胞中发挥着关键作用 流程。虽然RNA直接从DNA转录而来，但它们的序列、功能和翻译效率 都是通过转录后定位和编辑在细胞中主动调节的。这些过程是 对发育和细胞功能至关重要，在许多疾病中调节失调。尽管重要的是 对于转录后本地化和修饰，我们对时间的理解仍然存在重大差距， 机制，以及对这些事件的监管。据报道，有几种方法使研究人员能够 标记活细胞中的特定RNA并对其成像，或从细胞裂解产物下拉并对编辑后的转录本进行测序。 然而，仍然存在重大的技术限制，解决这些差距有望为 开发新的疗法和诊断学。我们之前已经开发并实施了新的 细胞中特定RNA的共价标记和成像方法以及A-R的下拉和浓缩 TO-I编辑了细胞RNA的转录本。我们计划的未来方向包括扩大这种化学品 生物学工具箱，为研究人员提供探测这些重要细胞的新的和改进的技术 流程。此外，最近的研究为不对称的假设提供了突破性的证据 RNA的本地化是一种调节编辑的机制。因此，拟议的一个重要重点是 研究是将我们的成像和编辑特定的下拉式方法结合起来，以促进对 这两个过程之间的相互作用。总而言之，这项研究旨在通过以下方式服务于科学界 提供对RNA本地化和修饰的监管以及开发和传播的见解 用于研究和调节生物系统的健壮且经过充分验证的技术。

Abstract RNA serves as the key intermediate for carrying genetic information and translating functional proteins, and numerous types of non-coding RNAs have also been recently discovered that play critical roles in cellular processes. While RNAs are directly transcribed from DNA, their sequence, function, and translation efficiency are all actively modulated in the cell though post-transcriptional localization and editing. These processes are essential for development and cellular function and are dysregulated in many diseases. Despite the importance of post-transcriptional localization and modification, significant gaps remain in our understanding of the timing, mechanisms, and regulation of these events. Several methods have been reported that enable researchers to label and image specific RNAs in living cells or pull-down and sequence edited transcripts from cell lysates. However, significant technological limitations still exist, and addressing these gaps holds promise for the development of new therapeutics and diagnostics. We have previously developed and implemented new methods for covalent labeling and imaging of specific RNAs in cells as well as pull-down and enrichment of A- to-I edited transcripts from cellular RNA. The future directions for our program include expanding this chemical biology toolbox to provide researchers with new and improved technologies for probing these important cellular processes. Additionally, recent studies have offered groundbreaking evidence for the hypothesis that asymmetric localization of RNA serves as a mechanism for regulating editing. Thus, a significant focus of the proposed research is to combine our imaging and editing-specific pull-down methods to advance understanding of the interplay between these two processes. Together, this research aims to serve the scientific community by providing insight into the regulation of RNA localization and modification as well as developing and disseminating robust and well-validated technologies for studying and modulating biological systems.