Abstract
Circular RNA (circRNA) are a recently discovered class of RNA characterized by a covalently-bonded back-splice junction. As circRNAs are inherently more stable than other RNA species, they may be detected extracellularly in peripheral biofluids and provide novel biomarkers. While circRNA have been identified previously in peripheral biofluids, there are few datasets for circRNA junctions from healthy controls. We collected 134 plasma and 114 urine samples from 54 healthy, male college athlete volunteers, and used RNASeq to determine circRNA content. The intersection of six bioinformatic tools identified 965 high-confidence, characteristic circRNA junctions in plasma and 72 in urine. Highly-expressed circRNA junctions were validated by qRT-PCR. Longitudinal samples were collected from a subset, demonstrating circRNA expression was stable over time. Lastly, the ratio of circular to linear transcripts was higher in plasma than urine. This study provides a valuable resource for characterization of circRNA in plasma and urine from healthy volunteers, one that can be developed and reassessed as researchers probe the circRNA contents of biofluids across physiological changes and disease states.
Highlights
Background and SummaryThe advent of next-generation sequencing has spurred the discovery of a growing list of RNA biotypes, many of which are detectable across species, detected in numerous biofluids, and have biological function
CircRNA were identified for a handful of mammalian genes[6,7,8]
CircRNAs are abundantly expressed in a number of human tissues and cell types, and circRNA expression changes during development, and as a response to extrinsic factors such as stress, immune response, and hormonal stimuli[12,13,14,15,16,17]. These endogenous RNAs are characterized by their circular structures, which are formed by a back-splicing event that covalently links the 3′ “tail” splice donor with the upstream 5′ splice acceptor “head” of the transcript, forming a back-spliced, or “head-to-tail” junction
Summary
The advent of next-generation sequencing has spurred the discovery of a growing list of RNA biotypes, many of which are detectable across species, detected in numerous biofluids, and have biological function. CircRNAs are abundantly expressed in a number of human tissues and cell types, and circRNA expression changes during development, and as a response to extrinsic factors such as stress, immune response, and hormonal stimuli[12,13,14,15,16,17] These endogenous RNAs are characterized by their circular structures, which are formed by a back-splicing event that covalently links the 3′ “tail” splice donor with the upstream 5′ splice acceptor “head” of the transcript, forming a back-spliced, or “head-to-tail” junction. Most studies of circRNA have small sample sizes or are based on targeted microarray data, rather than discovery-based methods This dataset includes more than 100 samples from 54 volunteers from two accessible biofluids (plasma and urine). While this might be a direct comparator for concussions, or other diseases more prevalent in young men, we expect this dataset to help begin to fill out a broader assessment of circRNAs present in healthy populations
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