An in vivo reporter system for measuring increased inclusion of exon 7 in @entity23247 mRNA: potential therapy of XXXX .

@entity1 @entity23247 @entity377 @entity179 @entity11280 @entity2057 @entity8870
@entity2057 ( @entity2057 ) is a @entity179 resulting from homozygous loss of the @entity11280 gene. @entity23247 , a nearly identical copy gene, is preserved in @entity2057 @entity1 . A single @entity377 difference between @entity11280 and @entity23247 causes exon 7 skipping in the majority of @entity23247 mRNA. Gene therapy through modulation of @entity23247 gene transcription in @entity2057 @entity1 may be possible. We constructed a series of @entity11280 mini-genes comprised of @entity11280 exon 6 to exon 8 sequences fused to green fluorescence protein (GFP) or luciferase reporters, to monitor @entity11280 exon 7 splicing. These reporters recapitulated the splicing patterns of the endogenous @entity11280 gene in stable cell lines. The @entity11280 -luciferase reporter was approximately 3.5-fold more active than @entity23247 -luciferase and @entity11280 -GFP intensities were visually distinguishable from @entity23247 -GFP. We have screened chemical inducers and inhibitors of kinase pathways using stable @entity11280 -reporter lines and found that the phosphatase inhibitor @entity8870 specifically stimulated exon 7 inclusion within @entity23247 mRNAs. This is the first compound identified that can stimulate exon 7 inclusion into transcripts derived from the endogenous @entity23247 gene. These results demonstrate that this system can be utilized to identify small molecules that regulate the splicing of @entity11280 exon 7.
