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

被引：37

作者：

Suzuki, Hitoshi ^{[1
]}

Aoki, Yoshitsugu ^{[2
]}

Kameyama, Toshiki ^{[3
]}

Saito, Takashi ^{[2
]}

Masuda, Satoru ^{[2
]}

Tanihata, Jun ^{[2
]}

Nagata, Tetsuya ^{[2
,4
]}

Mayeda, Akila ^{[3
]}

Takeda, Shin'ichi ^{[2
]}

Tsukahara, Toshifumi ^{[1
]}

机构：

[1] Japan Adv Inst Sci & Technol, Sch Mat Sci, Nomi, Ishikawa 9231292, Japan

[2] NCNP, Natl Inst Neurosci, Dept Mol Therapy, Kodaira, Tokyo 1878502, Japan

[3] Fujita Hlth Univ, Inst Comprehens Med Sci, Div Gene Express Mech, Toyoake, Aichi 4701192, Japan

[4] Tokyo Med & Dent Univ, Grad Sch Med & Dent Sci, Bunkyo Ku, Tokyo 1130034, Japan

来源：

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES | 2016年 / 17卷 / 10期

基金：

日本学术振兴会;

关键词：

pre-mRNA splicing; DMD; circular RNA; multiple exon skipping; DUCHENNE MUSCULAR-DYSTROPHY; PRE-MESSENGER-RNA; CIRCULAR RNAS; LARGE INTRON; GENE; JUNCTIONS; DELETION;

D O I：

10.3390/ijms17101722

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页数：16