A quick, and robust, and unbiased platform for circular RNAs isolation, discovery and profiling. - The goal of this proposal is to develop and validate a novel platform for the enrichment of the complete repertoire of circular RNAs (circRNAs). Thousands of circRNAs have been reported since their discovery >40 years ago from human transcriptome. The importance of circRNAs can be gauged from the fact that they are broadly expressed in eukaryotic cells, show cell and tissue-type specificity; and their expression is often conserved across species. CircRNAs have been implicated in neurological diseases, innate immunity, hepatocellular carcinoma and lung fibrosis demonstrating their role in cellular physiological and pathological pathways. The present approaches are reported to be biased in enriching circRNAs repertoire. Given their importance in various diseases, there is an urgent and unmet need for efficient circRNA isolation technology. In Phase-I application, we proposed two specific aims to develop Quick-circRNA platform, validate and compare its efficacy with current methods. In aim-1 we will develop and establish Quick-circRNA platform in various conditions and cell types in obtaining uniform an unbiased enrichment of circRNAs. In aim-2 we will compare the efficacy of Quick-circRNA with known methods in various cellular conditions. We believe that phase-I proposal will provide a benchmark for phase-II, which can streamline the Quick-circRNA platform for enriching circRNAs with high reproducibility and efficiency. In the end, we will develop a quick, and robust, and unbiased platform for circular RNAs isolation, discovery and profiling.
