Gene therapeutic approach using mutation-adapted U1 snRNA to correct a @entity8099 splice defect in XXXX -derived cells.

@entity1 @entity227 @entity73247 @entity8105 @entity6391 @entity8099 @entity214
@entity6391 ( @entity6391 ) is a disease that primarily affects the peripheral retina and ultimately causes @entity227 . X-chromosomal forms of @entity6391 are frequently caused by mutations in the @entity6391 GTPase regulator ( @entity8099 ) gene. We show that the novel splice donor site ( @entity214 ) mutation @entity73247 in intron 10 of @entity8099 cosegregates with @entity6391 in a five-generation Caucasian family. The mutation causes in-frame skipping of exon 10 from @entity8099 transcripts in @entity1 -derived primary fibroblasts. To correct the splice defect, we developed a gene therapeutic approach using mutation-adapted U1 small nuclear RNA (U1). U1 is required for @entity214 recognition of pre-mRNAs and initiates the splice process. The mutation described herein interferes with the recognition of the @entity214 by U1. To overcome the deleterious effects of the mutation, we generated four U1 isoforms with increasing complementarity to the @entity214 . Lentiviral particles were used to transduce @entity1 -derived fibroblasts with these U1 variants. Full complementarity of U1 corrects the splice defect partially and increases recognition of the mutant @entity214 . The therapeutic effect is U1-concentration dependent as we show for endogenously expressed @entity8099 transcripts in @entity1 -derived cells. U1-based gene therapeutic approaches constitute promising technologies to treat @entity214 mutations in inherited diseases including @entity8105 .
