Identification and function of regulatory RNA in the phototropic model organism Synechocystis sp. PCC 6803

非编码RNA(NcRNAs)在生命的所有领域都是基因表达的序列特异性调节因子。在之前的资助期间，我们已经解开了模式蓝藻聚球藻中几乎完整的一套ncRNAs和反义RNAs。利用转录组测序、详细的微阵列分析、ncRNA基因的计算预测以及广泛的Northern和5‘TAP-RACE实验相结合，全面和系统地研究了PCC6803。这些分析证明了数百个这样的基因座的存在，这些基因座产生了潜在的调节RNA分子。我们对在RNA代谢中具有关键功能的几个最有希望的ncRNA基因和蛋白质(RNase III，RNase E/G，Hfq)进行了基因敲除突变，以发展集胞藻。将PCC 6803转化为分析蓝藻调控RNA的真实模型。在新的资助时期，我们将把重点转移到几个选定的ncRNAs、质粒编码的CRISPR-RNAs、蓝藻HfQ同源物和RNA降解途径的功能分析上。

Non-coding RNAs (ncRNAs) serve as sequence-specific regulators of gene expression in all domains of life. In the previous funding period we have unraveled the nearly complete set of ncRNAs and antisense RNAs in the model cyanobacterium Synechocystis sp. PCC 6803 in a comprehensive and systematical way using a combination of transcriptome sequencing, detailed microarray analysis, computational prediction of ncRNA genes, complemented by extensive Northern and 5’ TAP-RACE experiments. These analyses demonstrated the presence of several hundred such loci giving rise to potentially regulatory RNA molecules. We performed knock-out mutations of genes for several of the most promising ncRNA genes and proteins with key functions in the metabolism of RNA (RNase III, RNase E/G, Hfq), developing Synechocystis sp. PCC 6803 into a true model for the analysis of regulatory RNAs in cyanobacteria. In the new funding period we will shift our focus on the functional analysis of several selected ncRNAs, plasmid-encoded CRISPR-RNAs, the cyanobacterial Hfq homologue and RNA degradation pathways.