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In this work, we used locked nucleic acid (LNA)–based SSOs to modulate splicing of TNF receptor 2 (TNFR2) pre-mRNA.
Antisense oligonucleotides (AON) have been reported to modulate splicing of pre-mRNA transcribed from mutated genes and to restore a normal reading frame in several diseases.
Of particular interest, the notion that SF2/ASF was first discovered as a cell type-specific regulator of another well-studied member of the polyomavirus family, SV40, based on its ability to modulate splicing of the viral early gene, thus effecting the expression of large T-antigen and small t-antigen expression at the post transcriptional level [17], [18].
Furthermore, the alternative splicing pattern of transcripts of several other SR genes was altered in a mutant, sr45-1, suggesthat thet the observed phenotypic abnormalities in sr45-1 are likely due to altered levels of SR protein isoforms, which in turn modulate splicing of other pre-mRNAs.
The ability to persistently modulate splicing of a target gene using ASOs provides a powerful method to model and characterize diseases in animals.
A common G to A polymorphism at codon 242 on exon 4 in the CCND1 gene has been shown to modulate splicing of the CCND1 transcript (Betticher et al, 1995).
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As MeCP2 is known to recruit HDACs to target genes for transcriptional repression, we tested whether MeCP2 modulates splicing of ASEs via recruiting the activities of HDACs.
19 The restoration of normal splicing by suppressing cryptic splicing, therefore, represents the very earliest manifestation of ASOs being used to modulate splicing for therapy.
The annealing of oligonucleotides directly complementary to splice site sequences has been used to modulate splicing decisions of different pre-mRNAs including tau, dystrophin and the apoptotic regulator Bcl-x [reviewed in [ 14]].
Our results raise the possibility that PUF60 and U2AF65 may belong to a family of factors that can modulate splicing based on substrate-specific, early recognition of distinct 3' splice sites.
A growing list of pharmacological agents that can modulate splicing is now emerging, with some demonstrating anti-tumor activity [ 46- 48].
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