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

用于 RNA 转录后修饰和捕获的化学生物学工具箱

基本信息

批准号：
10604335
负责人：
Jennifer Margaret Heemstra
金额：
$ 44.41万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-01 至 2027-04-30
项目状态：
未结题

项目摘要

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.

抽象的 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，许多类型的非编码RNA在细胞中起关键作用过程。虽然RNA直接从DNA转录，但它们的序列，功能和翻译效率通过转录后定位和编辑，都在单元格中进行了积极调制。这些过程是对于发育和细胞功能至关重要，在许多疾病中失调。尽管很重要在转录后定位和修改中，我们对时间安排的理解仍然存在很大的差距，这些事件的机制和调节。已经报道了几种方法，使研究人员能够活细胞中的标签和图像特异性RNA，或下拉和序列编辑的转录本来自细胞裂解物。但是，仍然存在重大的技术局限性，解决这些差距对开发新的治疗疗法和诊断。我们以前已经开发并实施了新的细胞中特定RNA的共价标记和成像的方法，以及下拉和富集A- TO-I从细胞RNA编辑的转录本。我们计划的未来指示包括扩展这种化学物质生物学工具箱为研究人员提供新的和改进的技术，以探测这些重要的细胞过程。此外，最近的研究为不对称的假设提供了开创性的证据 RNA的定位是调节编辑的机制。因此，提议的重点是研究是结合我们的成像和特定于编辑的下拉方法，以促进对这两个过程之间的相互作用。这项研究旨在通过提供有关RNA定位和修改的调节以及开发和传播的见解可用于研究和调节生物系统的强大且良好的技术。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Harnessing Nature's Molecular Recognition Capabilities to Map and Study RNA Modifications.

DOI：
10.1021/acs.accounts.2c00287
发表时间：
2022-08-16
期刊：
ACCOUNTS OF CHEMICAL RESEARCH
影响因子：
18.3
作者：
Felix, Ansley S.;Quillin, Alexandria L.;Mousavi, Shikufa;Heemstra, Jennifer M.
通讯作者：
Heemstra, Jennifer M.

Fluorescence Quenching of Xanthene Dyes during Amide Bond Formation Using DMTMM.