[HTML][HTML] Nuclear factor TDP‐43 and SR proteins promote in vitro and in vivo CFTR exon 9 skipping

E Buratti, T Dörk, E Zuccato, F Pagani… - The EMBO …, 2001 - embopress.org
E Buratti, T Dörk, E Zuccato, F Pagani, M Romano, FE Baralle
The EMBO journal, 2001embopress.org
Alternative splicing of human cystic fibrosis transmembrane conductance regulator (CFTR)
exon 9 is regulated by a combination of cis‐acting elements distributed through the exon
and both flanking introns (IVS8 and IVS9). Several studies have identified in the IVS8 intron
3′ splice site a regulatory element that is composed of a polymorphic (TG) m (T) n repeated
sequence. At present, no cellular factors have been identified that recognize this element.
We have identified TDP‐43, a nuclear protein not previously described to bind RNA, as the …
Alternative splicing of human cystic fibrosis transmembrane conductance regulator (CFTR) exon 9 is regulated by a combination of cis‐acting elements distributed through the exon and both flanking introns (IVS8 and IVS9). Several studies have identified in the IVS8 intron 3′ splice site a regulatory element that is composed of a polymorphic (TG) m (T) n repeated sequence. At present, no cellular factors have been identified that recognize this element. We have identified TDP‐43, a nuclear protein not previously described to bind RNA, as the factor binding specifically to the (TG) m sequence. Transient TDP‐43 overexpression in Hep3B cells results in an increase in exon 9 skipping. This effect is more pronounced with concomitant overexpression of SR proteins. Antisense inhibition of endogenous TDP‐43 expression results in increased inclusion of exon 9, providing a new therapeutic target to correct aberrant splicing of exon 9 in CF patients. The clinical and biological relevance of this finding in vivo is demonstrated by our characterization of a CF patient carrying a TG10T9 (ΔF508)/TG13T3 (wt) genotype leading to a disease‐causing high proportion of exon 9 skipping.
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