Endogenous Multiple Exon Skipping and Back-Splicing at the DMD Mutation Hotspot.
Endogenous Multiple Exon Skipping and Back-Splicing at the DMD Mutation Hotspot.
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DOI：
10.3390/ijms17101722
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发表时间：
2016-10-13
影响因子：
5.6
通讯作者：
Tsukahara T
中科院分区：
文献类型：
--
作者：
Suzuki H;Aoki Y;Kameyama T;Saito T;Masuda S;Tanihata J;Nagata T;Mayeda A;Takeda S;Tsukahara T
杜氏肌营养不良症(DMD)是一种严重的肌肉疾病。据报道，针对DMD基因外显子45-55的多外显子跳过(MES)可能会改善患者的症状，因为所有这些外显子的基因组缺失的患者表现出非常轻微的症状。因此，外显子45-55跳过治疗DMD已被认为是一种潜在的临床治疗方法。在此，我们用逆转录聚合酶链式反应(RT-PCR)检测了DMD内源性外显子44-56连接的mRNA转录本的表达，并鉴定了热点周围共有8种类型的MES产物。令人惊讶的是，最近报道的转录后内含子的5‘剪接位点(共转录剪接后剩余的内含子)作为Mess的剪接供体位点。我们还测试了外显子组合来产生DMD环状RNA(CircRNAs)，并确定了反向剪接的优先剪接位点，这些剪接位点不仅参与CircRNA的产生，而且参与Mess。我们的结果符合目前的CircRNA生成模型，表明上游转录后内含子触发MES并生成CircRNA，因为它的存在对于内含子内相互作用或极远端剪接至关重要。
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.
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影响因子：
14.9
作者：
SHAPIRO, MB;SENAPATHY, P
通讯作者：
SENAPATHY, P
影响因子：
4.8
作者：
COCQUERELLE, C;MASCREZ, B;BAILLEUL, B
通讯作者：
BAILLEUL, B
影响因子：
2
作者：
Nakamura, Akinorl;Yoshida, Kunihiro;Ikeda, Shu-ichi
通讯作者：
Ikeda, Shu-ichi
影响因子：
46.9
作者：
Jeck, William R.;Sharpless, Norman E.
通讯作者：
Sharpless, Norman E.
影响因子：
5.8
作者：
Koressaar, Triinu;Remm, Maido
通讯作者：
Remm, Maido