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PLoS One. 2016 Oct 13;11(10):e0164085. doi: 10.1371/journal.pone.0164085. eCollection 2016.

Dynamic ASXL1 Exon Skipping and Alternative Circular Splicing in Single Human Cells.

Author information

1
Departments of Bioengineering and Applied Physics, Stanford University, Stanford, CA, 94305, United States of America.
2
Departments of Oncology and Computational Biology, St. Jude Children's Research Hospital, Memphis, TN, 38105, United States of America.
3
Department of Biochemistry and Howard Hughes Medical Institute, Stanford University, Stanford, CA, 94305, United States of America.

Abstract

Circular RNAs comprise a poorly understood new class of noncoding RNA. In this study, we used a combination of targeted deletion, high-resolution splicing detection, and single-cell sequencing to deeply probe ASXL1 circular splicing. We found that efficient circular splicing required the canonical transcriptional start site and inverted AluSx elements. Sequencing-based interrogation of isoforms after ASXL1 overexpression identified promiscuous linear splicing between all exons, with the two most abundant non-canonical linear products skipping the exons that produced the circular isoforms. Single-cell sequencing revealed a strong preference for either the linear or circular ASXL1 isoforms in each cell, and found the predominant exon skipping product is frequently co-expressed with its reciprocal circular isoform. Finally, absolute quantification of ASXL1 isoforms confirmed our findings and suggests that standard methods overestimate circRNA abundance. Taken together, these data reveal a dynamic new view of circRNA genesis, providing additional framework for studying their roles in cellular biology.

PMID:
27736885
PMCID:
PMC5063410
DOI:
10.1371/journal.pone.0164085
[Indexed for MEDLINE]
Free PMC Article

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