Messenger RNA repair and restoration of protein function by spliceosome-mediated RNA trans-splicing

The functional repertoire of the human genome is amplified by the different ial assortment of exons. Spliceosome-mediated RNA trans-splicing can mobili ze these packets of genetic information to reprogram mRNAs. In principle, t his process could repair defective transcripts in loss-of-function genetic disorders in humans. We developed a tractable lacZ repair system to serve a s a model for these genetic disorders. Targeted pre-trans-splicing RNA mole cules efficiently and specifically repaired mutated lacZ transcripts and re stored enzymatic activity in human cells. The development of this model con firms the potential for spliceosome-mediated RNA trans-splicing in genetic repairs and provides a powerful tool for rational design and in vitro evolu tion of pre-trans-splicing molecules.