Reprogramming genetic information at the RNA level: optimizing tools to address specific biological questions

在 RNA 水平上重新编程遗传信息：优化工具来解决特定的生物学问题

• 批准号: 404867268
• 负责人: Professorin Dr. F. Nina Papavasiliou, Ph.D.
• 金额: --
• 依托单位: Interfakultäres Institut für Biochemie (IFIB)
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/404867268?language=en
• 关键词: Reprogramming; genetic; information; RNA; level

RNA editing is a robust and nearly ubiquitous modification (deamination) of RNA. Compared to most other modifications it is straightforward to identify as it alters nucleic acid sequence in a defined fashion (e.g. adenosine to inosine and cytosine to uracil, catalyzed by ADARs or APOBECs, respectively). It occurs reproducibly and with high prevalence in transcripts derived from cells of the immune system and the nervous system but also in transcripts derived from tumor cells from lung, breast, colon and most other tissues. Significant effort has been expended over the last decade to catalog editing, especially in the context of cancer. These efforts have led to correlations between editing load and disease outcomes. However, causality of editing of specific transcripts with biological outcomes has not been demonstrated, due to lack of proper tools to efficiently recapitulate editing of subsets of transcripts within specific cells, and to address the functional consequences of such targeted editing. Here, we propose to use (and continue to optimize) tools that allow efficient and multi-targeted ADAR editing, to recapitulate specific (natively occurring) editing events on groups of transcripts, which we have recovered from cancer cells, and to address the functional consequences of aggregate editing for pathways defined by the edited transcripts. We also propose to generate novel tools to allow targeted APOBEC editing which we can also apply to address the role of dual editing (ie both ADAR and APOBEC) on transcripts and the pathways those define.This proposal falls under the umbrella of Specific Priority Programme “Chemical Biology of Native Nucleic Acid Modifications ”. It brings together two groups with complementary expertise: the first group has been at the forefront of tool development for efficient and precise ADAR targeting. The other group has spent considerable time cataloguing instances of editing (in healthy tissue and in cancer) and then attempting to understand their functional relevance. In addition to gaining an understanding on the consequences of RNA editing, the tools we propose here to build will allow the targeting of RNAs in non-native settings (e.g. coding regions) and this will allow the re-coding of amino acids for therapeutic purposes.

RNA编辑是RNA的一种强大且几乎无处不在的修饰（脱氨基）。与大多数其他修饰相比，由于其以确定的方式改变核酸序列（例如，腺苷变为肌苷和胞嘧啶变为尿嘧啶，分别由ADAR或APOBEC催化），因此可以直接鉴定。它可重复发生，并且在源自免疫系统和神经系统细胞的转录物中具有高流行率，而且在源自肺、乳腺、结肠和大多数其他组织的肿瘤细胞的转录物中也具有高流行率。在过去的十年中，已经花费了大量的努力来编辑目录，特别是在癌症的背景下。这些努力导致了编辑负荷和疾病结果之间的相关性。然而，编辑特定转录物与生物学结果的因果关系尚未得到证实，这是由于缺乏适当的工具来有效地概括特定细胞内转录物子集的编辑，并解决这种靶向编辑的功能后果。在这里，我们建议使用（并继续优化）工具，允许有效和多靶向的阿达尔编辑，概括特定的（天然发生的）编辑事件组的转录本，我们已经从癌细胞中恢复，并解决聚合编辑的功能后果定义的途径编辑的转录本。我们还建议生成新的工具，以允许有针对性的APOBEC编辑，我们也可以应用于解决双重编辑（即阿达尔和APOBEC）对转录本和这些定义的途径的作用。该建议福尔斯属于特定优先计划“天然核酸修饰的化学生物学“的保护伞。它汇集了两个具有互补专长的小组：第一个小组一直处于工具开发的最前沿，以实现有效和精确的阿达尔目标。另一组花了相当多的时间对编辑的实例进行编目（在健康组织和癌症中），然后试图了解它们的功能相关性。 除了了解RNA编辑的后果外，我们在这里提出的工具将允许在非天然环境（例如编码区）中靶向RNA，这将允许出于治疗目的重新编码氨基酸。