Analysis of aberrant pre-messenger RNA splicing resulting from mutations in ATP8B1 and efficient in vitro rescue by adapted U1 small nuclear RNA.

摘要: ATP8B1 deficiency is a severe autosomal recessive liver disease resulting from mutations in the gene characterized by continuous phenotypical spectrum intermittent (benign recurrent intrahepatic cholestasis; BRIC) to progressive familial cholestasis (PFIC). Current therapeutic options are insufficient, and elucidating molecular consequences of could lead personalized mutation-specific therapies. We investigated effect on pre-messenger RNA splicing 14 at exon-intron boundaries using an vitro minigene system. Eleven mutations, mostly associated with PFIC phenotype, resulted aberrant complete absence correctly spliced product. In contrast, three led partially correct were BRIC phenotype. These findings indicate inverse correlation between level product severity. Expression modified U1 small nuclear RNAs (snRNA) complementary splice donor sites strongly improved or completely rescued for several located donor, as well acceptor, sites. one case, we also evaluated exon-specific snRNAs that, targeting nonconserved intronic sequences, might reduce possible off-target events. Although very effective correcting exon skipping, they induced retention short downstream intron. Conclusion: systematically found that majority total skipping. The amount inversely correlated Compensatory snRNAs, mutated sites, able improve definition efficiently be novel strategy other genetic diseases. (Hepatology 2015;61:1382-1391)