title: Messenger RNA repair and restoration of protein function by spliceosome-mediated RNA trans-splicing.
abstract: The functional repertoire of the human genome is amplified by the differential assortment of exons. Spliceosome-mediated RNA trans-splicing can mobilize these packets of genetic information to reprogram mRNAs. In principle, this process could repair defective transcripts in loss-of-function genetic disorders in humans. We developed a tractable lacZ repair system to serve as a model for these genetic disorders. Targeted pre-trans-splicing RNA molecules efficiently and specifically repaired mutated lacZ transcripts and restored enzymatic activity in human cells. The development of this model confirms the potential for spliceosome-mediated RNA trans-splicing in genetic repairs and provides a powerful tool for rational design and in vitro evolution of pre-trans-splicing molecules.
authors.affiliation: Intronn LLC, 840 Main Campus Drive, Raleigh, North Carolina 27606, USA.
authors.forename: M
authors.initials: M
authors.lastname: Puttaraju
authors.forename: J
authors.initials: J
authors.lastname: DiPasquale
authors.forename: C C
authors.initials: CC
authors.lastname: Baker
authors.forename: L G
authors.initials: LG
authors.lastname: Mitchell
authors.forename: M A
authors.initials: MA
authors.lastname: Garcia-Blanco
mesh_terms: D002458:Cell Fractionation; D002460:Cell Line; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D006801:Humans; D015151:Immunoblotting; D007763:Lac Operon; D016133:Polymerase Chain Reaction / Q000379:methods; D012326:RNA Splicing / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D017381:Spliceosomes / Q000378:metabolism*; D014162:Transfection; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: C505032:CFTR protein, human; D012333:RNA, Messenger; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D001616:beta-Galactosidase