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Endogenous Multiple Exon Skipping and Back-Splicing at the DMD Mutation Hotspot

Authors :
Toshifumi Tsukahara
Tetsuya Nagata
Shin'ichi Takeda
Akila Mayeda
Toshiki Kameyama
Jun Tanihata
Yoshitsugu Aoki
Takashi Saito
Satoru Masuda
Hitoshi Suzuki
Source :
International Journal of Molecular Sciences, Vol 17, Iss 10, p 1722 (2016), International Journal of Molecular Sciences, International Journal of Molecular Sciences; Volume 17; Issue 10; Pages: 1722
Publication Year :
2016
Publisher :
MDPI AG, 2016.

Abstract

Duchenne muscular dystrophy (DMD) is a severe muscular disorder. It was reported that multiple exon skipping (MES), targeting exon 45–55 of the DMD gene, might improve patients’ symptoms because patients who have a genomic deletion of all these exons showed very mild symptoms. Thus, exon 45–55 skipping treatments for DMD have been proposed as a potential clinical cure. Herein, we detected the expression of endogenous exons 44–56 connected mRNA transcript of the DMD using total RNAs derived from human normal skeletal muscle by reverse transcription polymerase chain reaction (RT-PCR), and identified a total of eight types of MES products around the hotspot. Surprisingly, the 5′ splice sites of recently reported post-transcriptional introns (remaining introns after co-transcriptional splicing) act as splicing donor sites for MESs. We also tested exon combinations to generate DMD circular RNAs (circRNAs) and determined the preferential splice sites of back-splicing, which are involved not only in circRNA generation, but also in MESs. Our results fit the current circRNA-generation model, suggesting that upstream post-transcriptional introns trigger MES and generate circRNA because its existence is critical for the intra-intronic interaction or for extremely distal splicing.

Details

ISSN :
14220067
Volume :
17
Database :
OpenAIRE
Journal :
International Journal of Molecular Sciences
Accession number :
edsair.doi.dedup.....7aad374883b67767233db07e0a693610