Characterization of post-transcriptional regulation of gene expression in heart diseases

信使rna结合蛋白（mrna -binding protein, mrbp）是调节选择性剪接和microRNAs的非编码rna家族，长度约为22个核苷酸，是心脏转录后基因调控的重要组成部分。它们功能的改变会导致心律失常、心肌病或心脏肥厚。例如CELF和MBNL家族的mrbp以及microrna miR-1和miR-208。全面了解它们的细胞特异性功能对于更好地理解它们在心脏病中的作用是必要的，并将有助于开发新的治疗方法。mrbp和microrna通过结合短核苷酸序列来识别它们的mRNA靶标，例如一些mrbp的CUG或CCUG和microrna的5 '近端“种子”区（前6至8个核苷酸）。由于难以可靠地鉴定mRNA靶标，对其生物学功能的了解受到限制。除了深度测序方法外，本项目还使用改进的交联和免疫沉淀方案，旨在鉴定心脏mrbp和microrna的mrna靶点，并旨在在转录组水平上表征其rna结合基序。它们的基因组序列变异及其在正常和病变心脏组织中的表达的关联将在一些心脏疾病中进行研究。

Messenger RNA-binding proteins (mRBPs) regulating alternative splicing and microRNAs, a family of non-coding RNAs of about 22 nucleotides in length, are important components of posttranscriptional gene regulation in the heart. An alteration in their function leads to arrhythmias, cardiomyopathies or cardiac hypertrophy. Examples are mRBPs of the CELF and MBNL family and the microRNAs miR-1 and miR-208. A thorough knowledge of their cell specific function is necessary for a better understanding of their role in cardiac diseases and will aid in the development of new therapies. MRBPs and microRNAs recognize their mRNA targets by binding to short nucleotide sequences, e.g. CUG or CCUG for some mRPBs and the 5‘ proximal “seed” region (the first 6 to 8 nucleotides) in the case of microRNAs. The understanding of their biological function has been limited by difficulties to reliably identify mRNA targets. Using a modified cross-linking and immunoprecipitation protocol in addition to deep sequencing methods this project aims to identify mRNA-targets of cardiac mRBPs and microRNAs and aims to characterize their RNA-binding motifs on a transcriptome wide level. The association of their genomic sequence variation and their expression in normal and diseased cardiac tissue will be investigated in a number of cardiac diseases.