UMMS Affiliation

Department of Medicine (LRB 326)

Publication Date

2-2-2006

Document Type

Article

Subjects

Alternative Splicing; Animals; Base Sequence; Cell Line; Cyclic AMP Response Element-Binding Protein; DNA; Exons; Humans; Introns; Mice; Models, Genetic; Molecular Sequence Data; Muscular Atrophy, Spinal; Nerve Tissue Proteins; Oligoribonucleotides, Antisense; *RNA Splicing; RNA-Binding Proteins; Sequence Deletion; Sequence Homology, Nucleic Acid; *Silencer Elements, Transcriptional

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

Humans have two nearly identical copies of the Survival Motor Neuron (SMN) gene, SMN1 and SMN2. In spinal muscular atrophy (SMA), SMN2 is not able to compensate for the loss of SMN1 due to exclusion of exon 7. Here we describe a novel inhibitory element located immediately downstream of the 5' splice site in intron 7. We call this element intronic splicing silencer N1 (ISS-N1). Deletion of ISS-N1 promoted exon 7 inclusion in mRNAs derived from the SMN2 minigene. Underlining the dominant role of ISS-N1 in exon 7 skipping, abrogation of a number of positive cis elements was tolerated when ISS-N1 was deleted. Confirming the silencer function of ISS-N1, an antisense oligonucleotide against ISS-N1 restored exon 7 inclusion in mRNAs derived from the SMN2 minigene or from endogenous SMN2. Consistently, this oligonucleotide increased the levels of SMN protein in SMA patient-derived cells that carry only the SMN2 gene. Our findings underscore for the first time the profound impact of an evolutionarily nonconserved intronic element on SMN2 exon 7 splicing. Considering that oligonucleotides annealing to intronic sequences do not interfere with exon-junction complex formation or mRNA transport and translation, ISS-N1 provides a very specific and efficient therapeutic target for antisense oligonucleotide-mediated correction of SMN2 splicing in SMA.

Rights and Permissions

Citation: Mol Cell Biol. 2006 Feb;26(4):1333-46. Link to article on publisher's site

DOI of Published Version

10.1128/MCB.26.4.1333-1346.2006

Related Resources

Link to Article in PubMed

Journal/Book/Conference Title

Molecular and cellular biology

PubMed ID

16449646

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