Open AccessDynamic ASXL1 Exon Skipping and Alternative Circular Splicing in Single Human Cells
Author(s) -
Winston Koh,
Veronica D. Gonzalez,
Sivaraman Natarajan,
Caroline Robert,
Patrick O. Brown,
Charles Gawad
Publication year - 2016
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0164085
Subject(s) - exon , circular rna , alternative splicing , exon skipping , rna splicing , biology , gene isoform , genetics , exonic splicing enhancer , computational biology , intron , rna , gene , microbiology and biotechnology
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.