Trans-Splicing in Drosophila

果蝇中的反式剪接

• 批准号: 8241030
• 负责人: Brenton R. Graveley
• 金额: $ 43.56万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8241030
• 关键词: Architecture; Biochemical; Biochemical Genetics; Bioinformatics; Biological; Biological Process; Caenorhabditis elegans; Cells; Cellular biology; Code; Complement; Culicidae; Data; Detection; Development; Drosophila genus; Drosophila melanogaster; Elements; Eukaryota; Event; Exons; Gene Expression; Genes; Genetic; Genomics; Goals; Health; Human; Hybrids; Introns; Joining Exons; Length; Link; Malignant Neoplasms; Maps; Messenger RNA; Mutation; Nematoda; Noise; Organism; Planarians; Process; Proteins; RNA Splicing; RNA, Messenger, Splicing; Reading; Spices; Spliced Genes; Spliced Leader RNA; Spliced Leader Sequences; Staging; Surveys; Testing; Trans-Splicing; Translating; Trypanosoma; Uncertainty; abstracting; design; genome sequencing; human disease; improved; insight; mRNA Precursor; public health relevance; research study

DESCRIPTION (provided by applicant): Abstract: Pre-messenger RNA (pre-mRNA) splicing is a process in which introns are removed to join exons together. This process is used during the synthesis of nearly all metazoan mRNAs (~93% of human mRNAs) and is an important means of regulating gene expression. With only a handful of exceptions, splicing occurs in cis - the exons that are joined together are located on the same pre-mRNA. In some eukaryotes, including nematodes, trypanosomes and planarians, splicing can occur in trans. In these cases, a specialized spliced leader RNA is spliced to the 5' end of protein coding RNAs. Interestingly, there are a few cases where a distinct type of trans-splicing has been shown to occur, namely, the splicing of exons from protein coding genes. The two best characterized examples are the mod(mdg4) and lola genes from Drosophila. In each of these genes, a group of common 5' exons can be spliced in trans to one of several (26 for mod(mdg4) or 22 for lola) variable 3' exons. One convincing case of trans-splicing has been shown to occur in mosquitos and one has been recently shown in humans. In each of these cases, trans-splicing of these genes was discovered fortuitously. As a result, the true extent of trans-splicing is unknown. Moreover, nothing is known about the mechanisms involved in trans-splicing. We have recently used deep sequencing to identify 80 new genes that are trans-spliced in Drosophila. The long- term goals of this project are to further explore the trans-splicing landscape in Drosophila and to determine the mechanisms involved in this process. We will first perform an exhaustive survey for trans-spliced genes throughout development in different Drosophila species. Second, we will perform experiments to test whether the trans-spliced mRNAs are translated and are functional. Finally, these experiments will be complemented by a combination of biochemical, genetic, genomic, cell biology, and bioinformatics experiments designed to determine the mechanisms by which trans- splicing occurs. Together these experiments will provide tremendous insight into the mechanisms of trans-splicing, a completely understudied yet important process. Given the potential utility of trans-splicing in treating human diseases and that trans-spliced mRNAs in humans have recently been linked to cancer, it is likely the discoveries we make will be of direct relevance to human health. PUBLIC HEALTH RELEVANCE: These experiments will provide tremendous insight into the mechanisms of trans- splicing. Given the potential utility of trans-splicing in treating human diseases and that trans-spliced mRNAs in humans have recently been linked to cancer, it is likely the discoveries we make will be of direct relevance to human health.

描述（由申请人提供）： 摘要：前信使RNA（pre-mRNA）剪接是去除内含子以将外显子连接在一起的过程。该过程用于几乎所有后生动物 mRNA（约 93% 的人类 mRNA）的合成，是调节基因表达的重要手段。除了少数例外，剪接发生在顺式中——连接在一起的外显子位于相同的前 mRNA 上。在一些真核生物中，包括线虫、锥虫和涡虫，剪接可以发生反式。在这些情况下，专门剪接的前导 RNA 被剪接到蛋白质编码 RNA 的 5' 端。有趣的是，在一些情况下，已证明发生了不同类型的反式剪接，即蛋白质编码基因的外显子剪接。两个最具特征性的例子是来自果蝇的 mod(mdg4) 和 lola 基因。在这些基因中的每一个中，一组常见的 5' 外显子可以反式剪接到几个（mod(mdg4) 为 26 个或 lola 为 22 个）可变 3' 外显子之一。一种令人信服的反式剪接案例已被证明发生在蚊子中，最近也有一种发生在人类身上。在每一个案例中，这些基因的转拼都是偶然发现的。因此，反式剪接的真实程度是未知的。此外，对于反式剪接所涉及的机制一无所知。我们最近使用深度测序鉴定了果蝇中 80 个反式拼接的新基因。该项目的长期目标是进一步探索果蝇的反式剪接景观并确定该过程中涉及的机制。我们将首先对不同果蝇物种整个发育过程中的反式剪接基因进行详尽的调查。其次，我们将进行实验来测试转剪的 mRNA 是否被翻译并具有功能。最后，这些实验将得到生化、遗传学、基因组、细胞生物学和生物信息学实验的组合的补充，旨在确定转拼发生的机制。这些实验将共同提供对反式剪接机制的深刻见解，这是一个完全未被充分研究但非常重要的过程。鉴于反式剪接在治疗人类疾病方面的潜在效用，以及人类反式剪接 mRNA 最近已与癌症相关，我们的发现很可能与人类健康直接相关。 公共卫生相关性： 这些实验将为转拼机制提供深入的见解。鉴于反式剪接在治疗人类疾病方面的潜在效用，以及人类反式剪接 mRNA 最近已与癌症相关，我们的发现很可能与人类健康直接相关。