基于三代高通量测序的环状RNA可翻译性研究

Circular RNA is a type of endogenous RNA widely paid attention in recent years, more and more circRNAs were found predicted by using NGS method. However, most of their functions are still unclear. Recent studies have been shown that partial circRNAs had the coding ability for proteins. Due to the expression of circRNA abundance is extremely low, less circRNA should be coded to protein. For such a trace amounts of circRNAs, ribosomes footprinting method, Ribo-seq and the short-reads based NGS method are still hard to large-scalely detect translatable circRNAs.Combining with the developed ribosome immunecoprecipitation technology, this proposal is aimed to build a highly efficient trace circRNA whole-length sequencing library preparation methods, to improve the circRNA whole-length database. Combined with different cells and different stress treatment, we hope to find the varations of the translatable circRNAs, and also the relationship with the mRNAs from the same host genes.This study will be helpful for circRNA function identification, early disease diagnostic and small molecules medicine screening.

环状RNA是近几年才被广泛关注的一类内源RNA，越来越多的circRNA被发现或预测，然而除了少数几种circRNA的功能被证实，大部分的circRNA生物学功能仍然未知。近期研究发现，除了具有miRNA海绵等内源性非编码RNA特性外，部分circRNA还具有蛋白质可编码性。由于circRNA本身的表达丰度极低，具有可编码性的circRNA量更少。对于如此微量circRNA，通过核糖体足迹法、Ribo-seq以及短reads二代测序等很难大规模鉴定可编码circRNA。本课题希望建立一种高效的微量circRNA全长大规模鉴定方法，用以完善并构建circRNA全长数据库，同时结合发展的核糖体免疫共沉淀技术和不同细胞系及不同应激处理，发现可编码circRNA的可翻译量的变化以及与对应的mRNA之间的关联性。本研究对于circRNA的功能鉴定、疾病早期诊断标及小分子药物筛选等具有重要的应用价值。

环状RNA是近几年才被广泛关注的一类内源RNA，越来越多的circRNA被发现或预测，然而除了少数几种circRNA的功能被证实，大部分的circRNA生物学功能仍然未知。目前被证明的circRNA功能主要包括miRNA海绵、蛋白质海绵、参与调控基因转录及可编码短肽等。然而要深入研究circRNA的这些功能，对于其完整的序列特征必须要准确的获取。目前基于短片段的二代测序加上基于拼接的生物信息学手段，在获取准确的全长circRNA序列信息方面仍然存在很多局限性。为此，本课题发展一种高效的基于长读长Nanopore测序平台的微量circRNA全长大规模鉴定方法，并通过基于片段重复的全长circRNA生物信息学分析方法，尽可能多的获得准确的circRNA全长信息，用以完善并构建circRNA全长数据库，并应用于其生物学功能的分析，包括其可编码性预测分析等。项目的成功实施，对于进一步研究circRNA在发育、疾病中的机制提供有效的手段，为circRNA在疾病诊断、创新药物研制、基础生物学研究等领域中的广泛应用提供基础性分析理论及使用工具。