Activation of a cryptic 5' splice site reverses the impact of pathogenic splice site mutations in the spinal muscular atrophy gene.
Activation of a cryptic 5' splice site reverses the impact of pathogenic splice site mutations in the spinal muscular atrophy gene.
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DOI：
10.1093/nar/gkx824
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发表时间：
2017-12-01
影响因子：
14.9
通讯作者：
Singh RN
中科院分区：
文献类型：
--
作者：
Singh NN;Del Rio-Malewski JB;Luo D;Ottesen EW;Howell MD;Singh RN
脊髓性肌萎缩症(SMA)是由存活运动神经元1(SMN1)基因的缺失或突变以及SMN2外显子7的主要跳跃引起的。目前唯一被批准的SMA治疗方法是针对位于外显子7 5‘剪接点(5’ss)下游的内含子剪接沉默蛋白N1(ISS-N1)的反义寡核苷酸。在这里，我们描述了一种通过激活隐蔽的5‘SS(CR1)来调控外显子7剪接的新方法。我们在SMN1的转录产物中发现了CR1的激活，SMN1在内含子7的第一位(G1C)携带致病的G-到-C突变。我们证明了CR1激活的工程U1单链RNA(EU1s)具有重新编程预mRNA剪接和恢复SMN1中的外显子7的独特能力，SMN1在外显子7的3‘s和5’s都带有广泛的致病突变。利用剪接耦合翻译报告，我们证明了由Eu1诱导的CR1激活产生的mRNAs产生全长的SMN。我们的发现强调了U1 SnRNP在剪接调控中的更广泛作用，并揭示了一种恢复SMN外显子7包涵体的新方法，为SMA的潜在治疗提供了可能。
Spinal muscular atrophy (SMA) is caused by deletions or mutations of the Survival Motor Neuron 1 (SMN1) gene coupled with predominant skipping of SMN2 exon 7. The only approved SMA treatment is an antisense oligonucleotide that targets the intronic splicing silencer N1 (ISS-N1), located downstream of the 5′ splice site (5′ss) of exon 7. Here, we describe a novel approach to exon 7 splicing modulation through activation of a cryptic 5′ss (Cr1). We discovered the activation of Cr1 in transcripts derived from SMN1 that carries a pathogenic G-to-C mutation at the first position (G1C) of intron 7. We show that Cr1-activating engineered U1 snRNAs (eU1s) have the unique ability to reprogram pre-mRNA splicing and restore exon 7 inclusion in SMN1 carrying a broad spectrum of pathogenic mutations at both the 3′ss and 5′ss of the exon 7. Employing a splicing-coupled translation reporter, we demonstrate that mRNAs generated by an eU1-induced activation of Cr1 produce full-length SMN. Our findings underscore a wider role for U1 snRNP in splicing regulation and reveal a novel approach for the restoration of SMN exon 7 inclusion for a potential therapy of SMA.
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影响因子：
--
作者：
Ahmad S;Bhatia K;Kannan A;Gangwani L
通讯作者：
Gangwani L
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
4.6
作者：
Howell MD;Ottesen EW;Singh NN;Anderson RL;Seo J;Sivanesan S;Whitley EM;Singh RN
通讯作者：
Singh RN
影响因子：
5.2
作者：
Divina, Petr;Kvitkovicova, Andrea;Vorechovsky, Igor
通讯作者：
Vorechovsky, Igor
影响因子：
12.4
作者：
Howell, Matthew D.;Ottesen, Eric W.;Singh, Ravindra N.
通讯作者：
Singh, Ravindra N.