Antisense suppression of donor splice site mutations in the dystrophin gene transcript

被引：12

作者：

Fletcher, Sue ^{[1
,2
]}

Meloni, Penny L. ^{[2
]}

Johnsen, Russell D. ^{[2
]}

Wong, Brenda L. ^{[3
]}

Muntoni, Francesco ^{[4
]}

Wilton, Stephen D. ^{[1
,2
]}

机构：

[1] Murdoch Univ, Ctr Comparat Genom, South St, Perth, WA 6150, Australia

[2] Univ Western Australia, Ctr Neuromuscular & Neurol Disorders, CNND M518,35 Stirling Highway, Crawley, WA 6009, Australia

[3] Univ Cincinnati Coll Med, Cincinnati Childrens Hosp Med Ctr, Dept Pediat, Cincinnati, OH 45229 USA

[4] Univ Coll London Inst Child Hlth London, Dubowitz Neuromuscular Ctr, London WC1N 1EH, England

来源：

MOLECULAR GENETICS & GENOMIC MEDICINE | 2013年 / 1卷 / 03期

基金：

英国医学研究理事会; 美国国家卫生研究院;

关键词：

Duchenne muscular dystrophy; exon skipping; splice mutation; splice-switching oligomer;

D O I：

10.1002/mgg3.19

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

We describe two donor splice site mutations, affecting dystrophin exons 16 and 45 that led to Duchenne muscular dystrophy (DMD), through catastrophic inactivation of the mRNA. These gene lesions unexpectedly resulted in the retention of the downstream introns, thereby increasing the length of the dystrophin mRNA by 20.2 and 36 kb, respectively. Splice-switching antisense oligomers targeted to exon 16 excised this in-frame exon and the following intron from the patient dystrophin transcript very efficiently in vitro, thereby restoring the reading frame and allowing synthesis of near-normal levels of a putatively functional dystrophin isoform. In contrast, targeting splice-switching oligomers to exon 45 in patient cells promoted only modest levels of an out-of-frame dystrophin transcript after transfection at high oligomer concentrations, whereas dual targeting of exons 44 and 45 or 45 and 46 resulted in more efficient exon skipping, with concomitant removal of intron 45. The splice site mutations reported here appear highly amenable to antisense oligomer intervention. We suggest that other splice site mutations may need to be evaluated for oligomer interventions on a case-by-case basis.

引用

页码：162 / 173

页数：12

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