RNA编辑影响RNA二级结构介导可变剪接的调控机制研究

Mutation-based rescue experiments indicate that combined mechanism based on the non-coding RNA secondary structure to direct the mutually exclusive splicing events occur in genes like 14-3-3ξ, Dscam and so on (Yang et al, Nat Struct Mol Biol, 2011). More interestingly, A-to-I RNA editing event was also identified in some of these RNA secondary structures; therefore it may affect the pairing pattern of the particular RNA duplex to achieve more spliced variants within the cell in a spatial and temporal manner. Based on this finding, by deletion and compensatory mutations, we will further investigate the mechanism how A-to-I RNA editing to direct novel RNA secondary structure formation. With the help of ADAR-/ADAR- mutant, RNAi and overexpression analysis, it would be much easier to understand the molecular mechanism of A-to-I RNA editing that affect mutually exclusive splicing events. It would be of great significance to clarify the regulatory mechanism of RNA secondary structure-mediated alternative splicing.

我们通过RNA补偿突变表明以非编码RNA二级结构基础的联合机制在14-3-3ξ, Dscam等基因的互斥可变剪接中的指导作用(Yang等, Nat Struct Mol Biol, 2011)。有趣的是，我们发现有些RNA二级结构发生A至I RNA编辑，从而影响RNA配对竞争而导致剪接体的比例发生改变。在此基础上，通过缺失和补偿突变技术，进一步分析A至I RNA编辑影响RNA二级结构竞争效应的动态，应用ADAR-/ADAR-突变体、RNAi和过度表达技术，深入分析RNA编辑对互斥可变剪接及相关性状的调控作用，探明A至I RNA编辑影响互斥剪接的分子机制，对从多层次阐明RNA二级结构介导的可变剪接调控具有重要意义。

RNA可变剪接和编辑是产生蛋白质多样性的两种重要途径，我们通过比较基因组学结合实验阐明了MRP1，Srp，Dscam等非编码RNA竞争二级结构指导互斥剪接的功能,发现在Srp， Dscam基因中外显子4、9发现存在进化保守的双向RNA二级结构，提出了双向竞争性RNA二级结构调控RNA互斥可变剪接的新模型，发现了可产生高度多样性的Dscam新基因和可变外显子选择机制的新模型，揭示了通过空间位阻和动态RNA结构联合调控Dscam 外显子簇17可变互斥剪接的机制，通过RNA-seq和比较分析等方法发现A至I RNA编辑位点，阐明RNA编辑通过二级结构介导与互斥可变剪接相互影响的可能分子机制；开展了用人工特异性RNA内切酶逆转相关基因的异常剪接来治疗1型强直性肌营养不良症研究，对从多层次理解转录后基因表达调控和以及疾病防治提供了线索和依据。部分研究工作已发表在Nat Commun,RNA 等国内外著名期刊上。培养硕士研究生4名、博士研究生4名，博士后2名。