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

• 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
• 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
• 来源: https://reporter.nih.gov/project-details/10604335
• 关键词: Address; Biology; Cell physiology; Cells; Chemicals; Communities; DNA; Development; Disease; Event; Future; Genetic Transcription; Image; Immune response; Label; Methods; Modification; Play; Post-Transcriptional RNA Processing; Process; Proteins; RNA; Regulation; Reporting; Research; Research Personnel; Role; Technology; Transcript; Translating; Translations; Untranslated RNA; biological systems; cell motility; genetic information; improved; insight; novel diagnostics; novel therapeutic intervention; novel therapeutics; posttranscriptional; programs; technology validation

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.

摘要 RNA是携带遗传信息和翻译功能蛋白质的关键中间体， 最近还发现了许多类型的非编码RNA，它们在细胞凋亡中起着关键作用。 流程.虽然RNA直接从DNA转录，但它们的序列、功能和翻译效率 均通过转录后定位和编辑在细胞中进行主动调节。这些过程 对于发育和细胞功能至关重要，并且在许多疾病中失调。尽管重要性 转录后定位和修饰，我们对时间的理解仍然存在重大差距， 机制和这些事件的调节。据报道，有几种方法使研究人员能够 标记和成像活细胞中的特异性RNA或来自细胞裂解物的下拉和序列编辑的转录物。 然而，仍然存在重大的技术限制，解决这些差距有望实现 开发新的治疗和诊断方法。我们已经开发并实施了新的 用于共价标记和成像细胞中的特异性RNA以及拉下和富集A- 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 流程.此外，最近的研究提供了开创性的证据，为假设，不对称 RNA的定位作为调节编辑的机制。因此，建议的一个重要重点是 研究是联合收割机结合我们的成像和编辑特定的下拉方法，以促进对 这两个过程之间的相互作用。这项研究旨在通过以下方式为科学界服务： 提供深入了解RNA定位和修饰的调节，以及开发和传播 用于研究和调节生物系统的强大且经过充分验证的技术。

项目成果

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.

• DOI: 10.1021/acsomega.2c03085
• 发表时间: 2022-09-20
• 期刊: ACS OMEGA
• 影响因子: 4.1
• 作者: Patterson, Kristen N.;Romero-Reyes, Misael A.;Heemstra, Jennifer M.
• 通讯作者: Heemstra, Jennifer M.