Development of a novel method for high-throughput discovery of trans- spliced RNAs in mammalian cells

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Wedevelopedanovelmethodforhigh-throughputdiscoveryoftrans-splicedRNAsinmammalian cells. Our method is based on a mate-pair analysis, which enables us to collect information of both ends of DNA fragments. In conventional mate-pair analysis, there must contain circularized DNAs derived from multiple DNAs and those mate-pair fragments can be a cause of false positive trans-spliced genes. To solve this problem we developed a novel method, which allows us to prepare circularized DNA libraries in which each circularized DNA is originated from each single DNA. By this method, we can make a systematic survey of the presence of trans-spliced RNAs in mammalian cells in a highly precise manner, and we could find out several candidate genes in hematopoietic stem cells. This method is useful for discovery of trans-spliced RNAs and will enable us to understand the role of trans-spliced RNAs in mammalian.