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

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 skipp...

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Autores principales: Suzuki, Hitoshi, Aoki, Yoshitsugu, Kameyama, Toshiki, Saito, Takashi, Masuda, Satoru, Tanihata, Jun, Nagata, Tetsuya, Mayeda, Akila, Takeda, Shin’ichi, Tsukahara, Toshifumi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5085753/
https://www.ncbi.nlm.nih.gov/pubmed/27754374
http://dx.doi.org/10.3390/ijms17101722
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author Suzuki, Hitoshi
Aoki, Yoshitsugu
Kameyama, Toshiki
Saito, Takashi
Masuda, Satoru
Tanihata, Jun
Nagata, Tetsuya
Mayeda, Akila
Takeda, Shin’ichi
Tsukahara, Toshifumi
author_facet Suzuki, Hitoshi
Aoki, Yoshitsugu
Kameyama, Toshiki
Saito, Takashi
Masuda, Satoru
Tanihata, Jun
Nagata, Tetsuya
Mayeda, Akila
Takeda, Shin’ichi
Tsukahara, Toshifumi
author_sort Suzuki, Hitoshi
collection PubMed
description 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.
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spelling pubmed-50857532016-11-01 Endogenous Multiple Exon Skipping and Back-Splicing at the DMD Mutation Hotspot Suzuki, Hitoshi Aoki, Yoshitsugu Kameyama, Toshiki Saito, Takashi Masuda, Satoru Tanihata, Jun Nagata, Tetsuya Mayeda, Akila Takeda, Shin’ichi Tsukahara, Toshifumi Int J Mol Sci Article 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. MDPI 2016-10-13 /pmc/articles/PMC5085753/ /pubmed/27754374 http://dx.doi.org/10.3390/ijms17101722 Text en © 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Suzuki, Hitoshi
Aoki, Yoshitsugu
Kameyama, Toshiki
Saito, Takashi
Masuda, Satoru
Tanihata, Jun
Nagata, Tetsuya
Mayeda, Akila
Takeda, Shin’ichi
Tsukahara, Toshifumi
Endogenous Multiple Exon Skipping and Back-Splicing at the DMD Mutation Hotspot
title Endogenous Multiple Exon Skipping and Back-Splicing at the DMD Mutation Hotspot
title_full Endogenous Multiple Exon Skipping and Back-Splicing at the DMD Mutation Hotspot
title_fullStr Endogenous Multiple Exon Skipping and Back-Splicing at the DMD Mutation Hotspot
title_full_unstemmed Endogenous Multiple Exon Skipping and Back-Splicing at the DMD Mutation Hotspot
title_short Endogenous Multiple Exon Skipping and Back-Splicing at the DMD Mutation Hotspot
title_sort endogenous multiple exon skipping and back-splicing at the dmd mutation hotspot
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5085753/
https://www.ncbi.nlm.nih.gov/pubmed/27754374
http://dx.doi.org/10.3390/ijms17101722
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