Analysis of RNA decay for alternatively spliced transcripts by RNA tagging

选择性剪接已被提出作为一种可能的机制，以增加蛋白质的数量显着相比，基因的数量。然而，人们一直在讨论它是否真的会产生功能性蛋白质产品。或者，它可以提供基因表达调控的转录后机制，或由剪接机制中的噪声引起。在这个项目中，我们的目标是研究选择性剪接使用一种新开发的方法，直接标记新转录的RNA，并确定RNA的半衰期与高精度。使用RNA标签和新型外显子阵列和RNA测序技术，可以确定单个外显子和可变剪接事件的RNA转录和衰变速率。为此，我们将开发用于确定可变剪接转录本的半衰期和识别剪接异构体之间的显著差异的方法。通过这种方式，我们的目标是定义一个预测模型来区分功能性剪接产物并表征相应蛋白质产物的调控。此外，将开发方法，以确定新的选择性剪接产物从RNA从头合成的测量和分析确定RNA衰变的机制。最后，不同RNA定量技术的比较将对每种方法的性能进行全面评估，并增加最终结果的置信度。

Alternative splicing has been proposed as a possible mechanism to increase the number of proteins significantly compared to the number of genes. However, it has been under discussion whether it actually results in functional protein products. Alternatively, it may provide a post-transcriptional mechanism for gene expression regulation or result from noise in the splicing machinery. In this project, we aim to investigate alternative splicing using a newly developed method for directly tagging newly transcribed RNA and determining RNA half-lives with high precision. Using RNA tagging and novel exon array and RNA sequencing technology, RNA transcription and decay rates can be determined for individual exons and alternative splicing events. For this purpose, we will develop methods for determining halflives of alternatively spliced transcripts and identifying significant differences between splicing isoforms. In this way, we aim to define a predictive model to distinguish functional splicing products and characterize the regulation of the corresponding protein products. Furthermore, methods will be developed to identify novel alternative splicing products from measurements of RNA de novo synthesis and to analyze the mechanisms determining RNA decay. Finally, the comparison of different RNA quantification techniques will provide a comprehensive assessment of the performance of each method and increase the confidence of the final results.